СПОСОБ ПОЛУЧЕНИЯ ИММОРТАЛИЗОВАННОЙ КЛЕТКИ ЧЕЛОВЕКА, СТАБИЛЬНО ТРАНСФИЦИРОВАННАЯ ИММОРТАЛИЗОВАННАЯ КЛЕТКА ЧЕЛОВЕКА, СПОСОБ РЕКОМБИНАНТНОЙ ПРОДУКЦИИ ЦЕЛЕВОГО БЕЛКА ЧЕЛОВЕКА, ПРИМЕНЕНИЕ ВЕКТОРА ТРАНСФЕКЦИИ

Изобретение относится к области биотехнологии, а именно к способу получения иммортализованной клетки человека, стабильно трансфицированной последовательностью нуклеиновой кислоты, стабильно трансфицированной иммортализованной клетке человека, полученной данным способом, способу рекомбинантной продукции целевого белка человека и применению вектора трансфекции. Способ получения иммортализованной клетки человека включает трансфекцию иммортализованной клетки-хозяина человека в бессывороточных условиях вектором трансфекции, содержащим ген, кодирующий целевой белок человека, промотор и сигнал полиаденилирования (полиА) гормона роста быка, где указанный промотор и сигнал полиА связаны с 5'- и 3'-концом гена, кодирующего целевой белок человека соответственно и начало репликации. Указанный вектор трансфекции дополнительно несет, по меньшей мере, один ген маркера селекции. Отбирают стабильно трансфицированные клетки. Предложенное изобретение позволяет получить стабильно трансфицированные иммортализованные ...

СЛИТЫЕ ПОЛИПЕПТИДЫ, СОДЕРЖАЩИЕ ДОМЕН WAP, И СПОСОБЫ ИХ ПРИМЕНЕНИЯ

Изобретение относится к области биотехнологии, конкретно к слитым белкам для ингибирования нейтрофильных серин-протеаз, и может быть использовано в медицине. Получают слитые белки имеющие по меньшей мере один полипептид человеческого секреторного ингибитора лейкоцитарных протеаз (SLPI), функционально связанный с полипептидом Fc-фрагмента иммуноглобулина, имеющего аминокислотную последовательность, которая по меньшей мере на 98% идентична аминокислотной последовательности SEQ ID NO: 10. Изобретение позволяет получить слитый полипептид, способный эффективно ингибировать активность нейтрофильных серин-протеаз и облегчать, таким образом, симптомы заболеваний или расстройств, связанных с избыточной экспрессией или активностью серин-протеаз у нуждающегося в этом субъекта. 5 н. и 10 з.п. ф-лы, 4 ил., 4 пр.

СЛИТЫЕ СЕРПИНОВЫЕ ПОЛИПЕПТИДЫ И СПОСОБЫ ИХ ПРИМЕНЕНИЯ

Настоящее изобретение относится к области иммунологии. Предложены выделенные слитые белки для ингибирования сериновых протеаз, а также их применения. Настоящее изобретение может найти дальнейшее применение в терапии различных заболеваний, в частности эмфиземы, псориаза и других. 6 н. и 12 з.п. ф-лы, 17 ил., 4 пр.

НОВЫЙ ВАРИАНТ АЛЬФА-1-АНТИТРИПСИНА, СПОСОБ ЕГО ПОЛУЧЕНИЯ И ПРИМЕНЕНИЯ

... 1. Вариант альфа-1-антитрипсина, полученный путем замены аминокислоты в определенном сайте между 1-м и 25-м положениями N-конца альфа-1-антитрипсина с целью добавления сайта гликозилирования.2. Вариант альфа-1-антитрипсина по п. 1, отличающийся тем, что вариант альфа-1-антитрипсина имеет от 1 до 3 добавленных к нему сайтов гликозилирования.3. Вариант альфа-1-антитрипсина по п. 1, отличающийся тем, что определенный сайт присутствует между 3-м и 13-м положениями N-конца.4. Вариант альфа-1-антитрипсина по п. 1, отличающийся тем, что определенный сайт присутствует в 9или 12положении N-конца.5. Вариант альфа-1-антитрипсина по п. 1, отличающийся тем, что определенные сайты присутствуют в 4-м и 9-м положениях, 4-м и 12-м положениях или 9-м и 12-м положениях.6. Способ получения варианта альфа-1-антитрипсина, включающий:a) замену аминокислоты в определенном сайте между 1-м и 25-м положениями N-конца альфа-1-антитрипсина с целью добавления сайта гликозилирования;b) культивирование клеток, трансформированных ...

Verfahren zur Isolierung von alpha-Antitrypsin

PROTEASE-INHIBITOREN DES ALPHA-1-ANTITRYPSIN TYPS MIT MODIFIZIERTER AKTIEVER STELLE UND DEREN HERSTELLUNG.

Verfahren zur Gewinnung von hochreinem, virusinaktiviertem „ 1 -Antitrypsin mittels Anionenaustauscher-Chromatographie

A process is disclosed for extracting highly purified, virus-inactivated alpha 1-anti-trypsin from a cryoprecipitate by means of anion exchanger chromatography with so-called "tentacular" materials. Membrane chromatography with appropriately modified surfaces may also be used in addition to or instead of conventional column chromatography.

Protein expression

RHEUMATOID ARTHRITUS TREATMENT

Thiol-active cysteinyl peptides for rheumatoid arthritis treatment

Protein variants and uses thereof

Methods of producing effective recombinant serine protease inhibitors and uses of these inhibitors.

A method of producing a recombinant serine protease inhibitor capable of effectively modulating serine protease activity is provided. Compositions capable of modulating serine protease activity and use of such compositions to regulate inflammatory processes in cells are also provided.

Methods of producing effective recombinant serine protease inhibitors and uses of these inhibitors

PROCEDURE FOR THE PRODUCTION OF A ALPHA-1 ANTITRYPSINLÖSUNG

ALPHA 1-ANTITRYPSIN-PRÄPARATION SOWIE VERFAHREN ZU DEREN HERSTELLUNG

PROCEDURE FOR THE PRODUCTION OF THE ALPHA-1-PROTEINASE OF INHIBITOR.

PROTEIN COMPLEXES WITH FACTOR VIII: C-ACTIVITY AND THEIR PRODUCTION

PROCEDURE FOR EXPRESSION OF ALPHA-1-ANTITRYPSIN IN BACTERIA AND ITS USE IN THERAPEUTIC COMPOSITIONS AS WELL AS VECTORS AND BACTERIA FUER SUCH A PROCEDURE AND THEIR PRODUCTION.

CLEANING OF ALPHA L PROTEINASEINHIBITOR MEANS NEW ONE CHROMATOGRAPHI SEPARATION CONDITIONS

Procedure for the isolation of the α1-Antitrypsins from human or animal body fluids

PROCEDURE FOR THE BLOKIEREN OF SECOND RECEPTORS

VARIANTS OF HUMAN ALPHA-1 ANTITRYPSIN

GLYCOLYTIC PROMTERS FOR REGULATED PROTEIN EXPRESSION PROTEASE INHIBITOR

Methods for purification of alpha-1-antitrypsin and apolipoprotein A-I

This invention relates to protein separation and purification methods for both alpha- 1-antitrypsin (AAT, also known as alpha-1 proteinase inhibitor, API, and A-Pl) and Apolipoprotein A-I (ApoA-l) from, for example, a fraction of human blood plasma. In certain embodiments, the invention provides methods for separating AAT from ApoA-l at the initial stage of purification, so that the same starting material can be used as a source for both proteins. The methods further pertain to providing compositions of AAT and of ApoA-l suitable for pharmaceutical use and are suitable for large-scale purification.

Pathogen-resistant monocot plants and method

Remodeling and glycoconjugation of peptides

Compositions, methods and uses for alpha-1 antitrypsin fusion molecules

Embodiments herein report compositions of and methods for making and using alpha-1 antitrypsin (AAT) fusion molecules or peptide derivatives thereof. In certain embodiments, compositions and methods relate to generating an AAT fusion molecule of use in pharmaceutically acceptable compositions to treat a subject in need of AAT therapy or treatment. In other embodiments, compositions and methods disclosed herein concern linking AAT or derivative thereof to an immune fragment. Fig. 7A Joint Scores following MSU-C16 Gouty Arthritis 1.256 p=0.032 l. j -, 0.25 - -r-- -r 'IV ,? <:,Q ...

Protein expression enhancing polypeptides

Fusion proteins comprising a protein expression enhancing polypeptide linked to a target protein binding domain and nucleic acid molecules encoding such fusion proteins are described for use in enhancing expression and/or location of a targeted protein of interest, for restoring lost functions in cells, and for treating disease. Additional fusion proteins comprising a target protein of interest modified with a fusion partner comprising a protein expression enhancing polypeptide are also disclosed.

LIPID NANOPARTICLE COMPOSITIONS AND METHODS FOR MRNA DELIVERY

Disclosed herein are compositions and methods for modulating the production of a protein in a target cell. The compositions and methods disclosed herein are capable of ameliorating diseases associated with protein or enzyme deficiencies.

LIPID NANOPARTICLE COMPOSITIONS AND METHODS FOR MRNA DELIVERY

Disclosed herein are compositions and methods for modulating the production of a protein in a target cell. The compositions and methods disclosed herein are capable of ameliorating diseases associated with protein or enzyme deficiencies.

Process for scaled-up production of recombinant proteins using transgenic plant suspension cultures

HIGH EXPRESSION OF FOREIGN GENES IN SCHIZOSACCHAROMYCES POMBE

DNA CONSTRUCT FOR IN VIVO EXPRESSION OF A HUMAN GENE

Alpha-1-antitrypsin and antithrombine-iii variants

METHOD FOR SEPARATING AND/OR ISOLATING PLASMA PROTEINS BY ANNULARCHROMATOGRAPHY

The invention relates to a method for separating and/or isolating especially human plasma proteins from a mixture containing plasma proteins. According to said method the mixture is deposited on an annular separating medium; the annular separating medium is rotated substantially vertically about an axis defined in the direction of flow of the mixture through the annular separating medium; an eluant is fed through the annular separating medium; and the fractions emerging at the end of the annular separating medium are collected.

METHOD, COMPOSITION, AND ARTICLE OF MANUFACTURE FOR PROVIDING ALPHA-1 ANTITRYPSIN

The present invention provides a method for providing alpha-1 antitrypsin ((.alpha.1-AT) to a subject, in particular a method for treating or preventing a disorder or disease associated with .alpha.1-AT deficiency in the subject, wherein the method comprises providing, subcutaneously, a therapeutically or prophylactically effective amount of .alpha.1-AT to the subject. Also provided is a composition and article of manufacture comprising .alpha.1-AT, in particular a formulation suitable for subcutaneous administration of .alpha.1-AT.

METHOD FOR PRODUCING PROTEINS IN PICHIA PASTORIS THAT LACK DETECTABLE CROSS BINDING ACTIVITY TO ANTIBODIES AGAINST HOST CELL ANTIGENS

Methods for producing proteins and glycoproteins in Pichia pastoris that lack detectable cross binding activity to antibodies made against host cell antigens are described. In particular, methods are described wherein recombinant Pichia pastoris strains that do not display a ß-mannosyltransferase 2 activity with respect to an N-glycan or O-glycan and do not display at least one activity selected from a ß-mannosyltransferase 1, 3, and 4 activity to produce recombinant proteins and glycoproteins. These recombinant Pichia pastoris strains can produce proteins and glycoproteins that lack detectable a-mannosidase resistant ß-mannose residues thereon and thus, lack cross binding activity to antibodies against host cell antigens. Further described are methods for producing bi-sialylated human erythropoietin in Pichia pastoris that lack detectable cross binding activity to antibodies against host cell antigens.

METHOD OF PREPARING ALPHA-1-PROTEINASE INHIBITOR

Inventors: MICHAEL A. SHEARER PAMELA K. SASAGAWA and RONALD H. HEIN Invention: IMPROVED METHOD OF PREPARING ALPHA-1-PROTEINASE INHIBITOR There is disclosed an improved method for separating one of alpha-1-proteinase inhibitor (also known as alpha-1 antitrypsin) and antithrombin-III from an aqueous solution of plasma proteins containing the same, such as from Cohn Fraction IV-1, Cohn Fraction IV, reworks of Cohn Fraction IV and IV-l, Cohn Effluent II \A III and Cohn Effluent I. The method includes the steps of first holding an aqueous solution of plasma proteins containing one of alpha-l-proteinase inhibitor and antithrombin-III in a relatively large volume of buffer solution as solvent and at a pH adjusted to be relatively basic when compared to conditions heretofore known, and at a temperature in the range of from 2 - 50.degree. C for a period of about 0.2 - 24 hours. Following the above treatment, alpha-1-proteinase inhibitor and antithrombin-III are obtained by applying conventional ...

COMPOSITIONS AND METHODS FOR TREATING ALPHA-1 ANTITRYPSIN DEFICIENCEY

Compositions and methods for expressing alpha 1 antitrypsin (AAT) in a host cell are provided. Also provided are compositions and methods for treating subjects having alpha 1 antitrypsin deficiency (AATD).

COMPOSITIONS AND METHODS FOR TREATING ALPHA-1 ANTITRYPSIN DEFICIENCY

The present invention features compositions and methods for editing deleterious mutations associated with alpha-1 anti-trypsin (A1AT) deficiency. In particular embodiments, the invention provides methods for correcting mutations in an A1AT polynucleotide using an adenosine deaminase base editor, ABE8, having unprecedented levels of efficiency.

OPTIMIZED PROMOTER SEQUENCES, INTRON-FREE EXPRESSION CONSTRUCTS AND METHODS OF USE

The invention provides expression cassettes. In certain embodiments, an expression cassette comprises (a) a regulatory element at least 90% identical to the sequence of any of SEQ ID NOs:2-67, and (b) a nucleic acid sequence encoding a Factor VIII protein having a B domain deletion (FVIII-BDD), where the nucleic acid sequence of (a) is at least 90% identical to the sequence of SEQ ID NO:77, where the regulatory element is operably linked to the nucleic acid sequence, and where no intron is present between the regulatory element and the nucleic acid sequence encoding FVIII-BDD, or where no more than 0 107 nucleotides of untranslated nucleic acid is between the regulatory element and the nucleic acid sequence encoding FVIII-BDD. In certain embodiments, expression cassettes contain sequence elements having CpG(s) substituted with CpT, CpA, TpG, or ApG at the same position(s) or has CpG reduced nucleic acid sequences.

VECTORS AND METHODS OF USE

This disclosure provides vectors and strategies for increasing the efficiency of gene therapy in hepatocytes. The efficiency of the delivery of a corrected gene or wild type gene is improved through the use of delivery to hepatocytes via intrahepatic (parenchyma) administration or administration via the portal vein. In addition, the corrected gene or wild type gene is delivered using an isolated exogenous nucleic acid comprising a promoter that is specifically expressed in hepatocytes. These methods and isolated exogenous nucleic acids are useful to correct gene defects in the liver such as inherited diseases of the liver.

PERFUSION COMPOSITIONS AND METHODS OF USING ALPHA-1 ANTI-TRYPSIN IN EX VIVO ORGAN PERFUSION

Perfusion solutions comprising AIAT for the ex vivo perfusion of donor organs are provided to improve donor organ quality and repair damaged donor organs for transplantation. Methods of ex vivo perfusion of donor organs with AlAT-containing perfusion solutions under normothermic temperatures are also provided.

MODIFIED SERPINS FOR THE TREATMENT OF BLEEDING DISORDERS

This invention relates pro-coagulant serpin molecules engineered by modification of the P4, P2, P1 and/or P1' residues within the reactive center loop (RCL) to display increased specificity for anticoagulant proteases. These modified serpin molecules may be useful in therapy, for example as pro-coagulants for the treatment of bleeding.

PURIFICATION OF CELL CULTURE DERIVED ALPHA1 PROTEASE INHIBITOR

Described herein are methods for purifying recombinant, cell culture derived alpha1-protease inhibitor and removing a colored species that co-purifies with the recA1PI protein. Also described are methods for reducing the iron in cell culture derived alpha1- protease inhibitor.

SERINE PROTEASE INHIBITOR (SERPIN) PEPTIDES AND METHODS THEREOF

Serpin peptides, and variants and derivatives thereof, are described herein. These peptides are unexpectedly potent anti-inflammatory agents and are therefore useful to treat autoimmune, inflammatory and metabolic diseases.

.ALPHA.-1-ANTICHYMOTRYPSIN ANALOGUES HAVING CHYMASE INHIBITING ACTIVITY

The invention provides analogues of .alpha.-1-antichymotrypsin having amino acid substitutions at positon 358. .alpha.-1-antichymotrypsin analogues having amino acid substitutions at positions 356-361 and analogues having amino acid substitutions at positions 356-361 wherein the amino acid at position 358 is substituted are also within the scope of the invention. These analogues exhibit chymase inhibitory activity. Also provided are novel .alpha.-1-antichymotrypsins having an N-terminal extension of methionine-alanine-serine or alanine-serine. Expression vectors for the production of .alpha.-1-antichymotrypsins are also provided. The present invention also provides host cells and cell cultures capable of expressing analogues of .alpha.-1-antichymotrypsins, as well as protein preparations from the host cells. Methods of producing and using the analogues of .alpha.-1-antichymotrypsins to inhibit chymase activity are also provided.

METHODS OF PRODUCING EFFECTIVE RECOMBINANT SERINE PROTEASE INHIBITORS AND USES OF THESE INHIBITORS

A method of producing a recombinant serine protease inhibitor capable of effectively modulating serine protease activity is provided. Compositions capable of modulating serine protease activity and use of such compositions to regulate inflammatory processes in cells are also provided.

PROCESS FOR SEPARATING ALPHA-1-PROTEINASE INHIBITOR FROM COHN FRACTION IV1 +IV4 PASTE

The present invention is directed to a process for purifying alpha-1-PI. The process comprises providing an impure protein fraction which comprises alpha-1-PI. The impure protein fraction is suspended in an aqueous solution at pH 6. Insoluble proteins are recovered and resuspended in aqueous solution at pH 8,5. PEG is added to precipitate .alpha.-2 proteins. To the PEG supernatant precipitation, which comprises alpha-1-PI, is added ZnCl2 to precipitate crude alpha-1-PI. The crude alpha-1-PI is resolubilized and apllied to an anion-exchange medium. A fraction comprising alpha-1-PI is recovered from the anion-exchang e medium. Alpha-1-PI purified by the process has a specific activity about 1.0 units/OD280.

Verfahren zur Isolierung des a1-Antitrypsins aus menschlichen oder tierischen Körperflüssigkeiten

Verfahren zur Isolierung des a1-Antitrypsins aus menschlichen oder tierischen Körperflüssigkeiten

Verfahren zur Isolierung von a1-Antitrypsin

Druckrohr mit Aussenverstärkung für Wasserkraftanlagen

PROCESS FOR THE PREPARATION OF PROTEASE INHIBITORS.

СПОСОБ РАЗДЕЛЕНИЯ И/ИЛИ ВЫДЕЛЕНИЯ ПРОТЕИНОВ ПЛАЗМЫ МЕТОДОМ КОЛЬЦЕВОЙ ХРОМАТОГРАФИИ

... 1. Способ разделения и/или выделения протеинов плазмы человека из плазмы крови или смеси, содержащей вирусно-инактивированные протеины плазмы, с использованием метода кольцевой хроматографии, отличающийся тем, что разделение осуществляют на разделяющей среде, имеющей слой аппликационной среды, представляющей собой сферические частицы с гидрофобной поверхностью. 2. Способ по п.1, отличающийся тем, что указанная смесь содержит, по меньшей мере, два протеина плазмы, подлежащих разделению. 3. Способ по п.1, где указанная разделяющая среда, имеющая кольцевую конфигурацию, используется для ионообменной, гель-проникающей, эксклюзионной по размеру молекул, или аффинной хроматографии, или для хроматографии, основанной на гидрофобных взаимодействиях. 4. Способ по п.1, где указанными протеинами плазмы являются ингибитор интер-α-трипсина, α1 -антитрипсин, антитромбин III, иммуноглобулины, такие как IgG, сывороточный альбумин человека или гликопротеины, предпочтительно из каскада свертывания, или витамин ...

METHOD OF PREPARATION Of a STOCK, IN PARTICULAR OF YEAST, PROCESSED BY a VECTOR Of EXPRESSION, WHICH CAN BE CULTIVEE ON a COMPLETE MEDIUM WITHOUT PRESSURE OF SELECTION AND STOCK THUS OBTAINED

VARIANTS [...]-TO-[...] GLYCOSYLATED HUMAN AND PRODUCING PROCESS

METHOD FOR OBTAINING A CONCENTRATE OF HAS 1-ANTITRYPSIN FROM HUMAN PLASMA AND ITS USE AS A DRUG

Process for the insulation of the alpha-antitrypsin starting from the organic liquids of human or animal origin

METHOD OF PREPARATION Of a STOCK, IN PARTICULAR OF YEAST, PROCESSED BY a VECTOR Of EXPRESSION, WHICH CAN BE CULTIVEE ON a COMPLETE MEDIUM WITHOUT PRESSURE OF SELECTION AND STOCK THUS OBTAINED

Recombinant glycoprotein with reduced antenna unary fucosylation

Optimized promoter sequences, intron-free expression constructs and methods of use

The invention provides expression cassettes. In certain embodiments, an expression cassette comprises (a) a regulatory element at least 90% identical to the sequence of any of SEQ ID NOs:2-67, and (b) a nucleic acid sequence encoding a Factor VIII protein having a B domain deletion (FVIII-BDD), where the nucleic acid sequence of (a) is at least 90% identical to the sequence of SEQ ID NO:77, where the regulatory element is operably linked to the nucleic acid sequence, and where no intron is present between the regulatory element and the nucleic acid sequence encoding FVIII-BDD, or where no more than 0-107 nucleotides of untranslated nucleic acid is between the regulatory element and the nucleic acid sequence encoding FVIII-BDD. In certain embodiments, expression cassettes contain sequence elements having CpG(s) substituted with CpT, CpA, TpG, or ApG at the same position(s) or has CpG reduced nucleic acid sequences.

PURIFICATION OF CELL CULTURE DERIVED ALPHA1 PROTEASE INHIBITOR

Described herein are methods for purifying recombinant, cell culturederived alphai-protease inhibitor and removing a colored species thatco-purifies with the recAlP1 protein. Also described are methods forreducing the iron in cell culture derived alpha-protease inhibitor. (NoSuitable Figure) ...

ALPHA 1-ANTITRYPSIN COMPOSITIONS AND TREATMENT METHODS USING SUCH COMPOSITIONS

LYSOSOMAL POLYPEPTIDES, METHODS OF MAKING AND USING

The invention generally relates to compositions and methods for producing lysosomal proteins that have altered glycan structure, such that the protein can be delivered efficiently into the lysosomes of target cells (such as macrophages). The lysosomal proteins are produced by modifying the glycosylation pathways in a host cell using an RNA effector molecule, such as an siRNA. Glycan-modified lysosomal proteins produced using the methods described herein have improved properties, including e.g., increased specific uptake by target cells (such as macrophages).

'alpha'-1-ANTICHYMOTRYPSIN ANALOGUES HAVING CHYMASE INHIBITING ACTIVITY

The invention provides analogues of 'alpha'-1-antichymotrypsin having amino acid substitutions at positon 358. 'alpha'-1-antichymotrypsin analogues having amino acid substitutions at positions 356-361 and analogues having amino acid substitutions at positions 356-361 wherein the amino acid at position 358 is substituted are also within the scope of the invention. These analogues exhibit chymase inhibitory activity. Also provided are novel 'alpha'-1-antichymotrypsins having an N-terminal extension of methionine-alanine-serine or alanine-serine. Expression vectors for the production of 'alpha'-1-antichymotrypsins are also provided. The present invention also provides host cells and cell cultures capable of expressing analogues of 'alpha'-1-antichymotrypsin, as well as protein preparations from the host cells. Methods of producing and using the analogues of 'alpha'-1-antichymotrypsin to inhibit chymase activity are also provided.

METHODS FOR PURIFICATION OF ALPHA-1-ANTITRYPSIN AND APOLIPOPROTEIN A-1

This invention relates to protein separation and purification methods for both alpha-1-antitrypsin (AAT, also known as alpha-1 proteinase inhibitor, API, and A.sub.1-PI) and Apolipoprotein A-I (ApoA-1) from, for example, a fraction of human blood plasma. In certain embodiments, the invention provides methods for separating AAT from ApoA-1 at the initial stage of purification, so that the same starting material can be used as a source for both proteins. The methods further pertain to providing compositions of AAT and of ApoA-1 suitable for pharmaceutical use and are suitable for large-scale purification.

METHODS AND COMPOSITIONS FOR SINGLE CHAIN VARIABLE REGION ENOX2 ANTIBODIES FOR CANCER DETECTION AND DIAGNOSIS

Cancers of different cellular or tissue origins express different ENOX2 cancer isoforms or combinations of isoforms and shed these proteins into the circulation. Herein are disclosed methods both for cancer detection and diagnosis of particular origin, based on the patterns and molecular weights of the isoforms which allow the identification of the cell type and or tissue of origin of the neoplasm. Relative ENOX2 amounts are proportional to tumor burden and provide a reliable measure of response to therapy and disease progression. Also provided is the amino acid sequence to which the scFv antibodies bind as the molecular basis for the specificity of the test. 1. A single chain variable region ENOX2 antibody linked to an alkaline phosphatase or other single step agent for detection or imaging.2. The antibody of claim 1 , wherein the single chain variable region ENOX2 antibody comprises the polypeptide sequence substantially as shown in SEQ ID NO:5.3. The antibody of claim 2 , wherein the antibody further comprises a detection aid.4. The antibody of claim 3 , wherein the detection aid is suitable for use in enzymatic claim 3 , chromagenic claim 3 , chemiluminescent claim 3 , magnetic or fluorescent methods.5. The antibody of claim 3 , wherein the single chain variable region ENOX2 antibody is linked to an S-tag.6. The antibody of claim 1 , wherein the single chain variable region ENOX2 antibody is coupled to a single step agent for detection or imaging.7. The antibody of claim 1 , wherein the single chain variable region ENOX2 antibody is chemically linked directly to an alkaline phosphatase.8. The antibody comprising the single chain variable region antibody of in aqueous solution at a concentration suitable for use as an assay reagent.9. A cell that expressed the antibody of .10. The cell of claim 9 , wherein the cell is a bacterium.11. A method for detecting cancer comprisingobtaining a sample from a patient suspected of comprising ENOX2 variants associated with a ...

Method for purification of alpha-1-antitrypsin

A streamlined method for purifying alpha-1-antitrypsin (AAT) from an AAT-containing protein mixture, such as a Cohn fraction IV precipitate, is provided. In the method of the invention, contaminating proteins are destabilized by cleavage of disulfide bonds with a reducing reagent, such as a dithiol, which does not affect AAT. The destabilized proteins are then preferentially adsorbed on a solid protein-adsorbing material, without the addition of a salt as a precipitant. Separation of the solid adsorbent from the solution leaves a purified AAT solution that is directly suitable for chromatographic purification, without the need for extensive desalting as in prior art processes. A process incorporating this method, which provides pharmaceutical grade AAT in high yield on a commercial scale, is also described.

MAMMALIAN ALVEOLAR MACROPHAGES DERIVED FROM PLURIPOTENT CELLS

Alveolar-like macrophages and a method for generating alveolar-like macrophages from hemangioblasts is provided. The method comprises the steps of: i) culturing the hemangioblasts in a hematopoietic-inducing medium comprising vascular endothelial growth factor (VEGF), stem cell factor (SCF) and interleukin-3 (IL-3) for a sufficient period of time to generate macrophages, and ii) culturing the macrophages in an alveolar macrophage-inducing medium comprising granulocyte macrophage colony stimulating factor (GM-CSF), and optionally macrophage colony stimulating factor (M-CSF), under suitable conditions and for a sufficient period of time to yield alveolar-like macrophages. 1. A population of isolated mammalian alveolar macrophages , wherein said alveolar macrophages:i) express CD11c;ii) do not express Myb;iii) are capable of being cultured for at least 1 month at a doubling rate of no more than about 5 days; andiv) are derived from pluripotent stem cells.2. The population of isolated mammalian alveolar macrophages as defined in claim 1 , wherein said alveolar macrophages exhibit a doubling rate of 3 days or less.3. The population of isolated mammalian alveolar macrophages as defined in claim 1 , wherein said alveolar macrophages are capable of being expanded for at least 1 year.4. The population of mammalian alveolar macrophages as defined in claim 1 , wherein said alveolar macrophages exhibit at least about 10% greater phagocytic activity as compared to primary alveolar macrophages.5. The population of isolated mammalian alveolar macrophages as defined in claim 1 , wherein said alveolar macrophages exhibit:i) at least 50% greater expression of CD11b than CD11b expression as compared to primary alveolar macrophages; orii) at least a 2-fold increase in expression of LSR or RUN×2 as compared to primary alveolar macrophages.6. The population of isolated mammalian alveolar macrophages of claim 1 , wherein said alveolar macrophages express interferon-gamma claim 1 , ...

COMPOSITIONS AND METHODS FOR TREATING ALPHA-1 ANTITRYPSIN DEFICIENCY

Compositions and methods for expressing alpha 1 antitrypsin (AAT) in a host cell are provided. Also provided are compositions and methods for treating subjects having alpha 1 antitrypsin deficiency (AATD).

DIAGNOSIS AND/OR PROGNOSIS OF PARKINSON'S DISEASE DEMENTIA

Remodelling and glycoconjugation of follicle-stimulating hormone (FSH)

The invention includes methods and compositions for remodeling a peptide molecule, including the addition or deletion of one or more glycosyl groups to a peptide, and/or the addition of a modifying group a peptide.

Remodelling and glycoconjugation of antibodies

The invention includes methods and compositions for remodeling a peptide molecule, including the addition or deletion of one or more glycosyl groups to a peptide, and/or the addition of a modifying group a peptide.

PERICARDIAL THERAPEUTIC AND DIAGNOSTIC AGENT DELIVERY

A method for treating a patient's heart which comprises delivering a gene therapy agent into the pericardial sac around the patient's heart. The agent is introduced surgically or by transvascular means such as a catheter which has been introduced percutaneously or otherwise. Introducing the gene therapy agent into the pericardial sac contains the agent, allowing high concentration of the agent adjacent large regions of the epicardium and pericardium without spillage or systemic distribution to other organs or tissues. The gene therapy agents of this invention comprise vectors for transferring genetic information to the epicardial cells in vivo or harvested cells which have been genetically engineered in vitro. In a preferred embodiment, a catheter is percutaneously introduced, such as through the femoral artery, and guided upstream into the left ventricle. The distal end of the catheter advanced until it penetrates through the epicardium so that agent can be introduced into the pericardial ...

PROTEIN COMPLEXES HAVING FACTOR VIII:C ACTIVITY AND PRODUCTION THEREOF

НОВЫЙ ВАРИАНТ АЛЬФА-1-АНТИТРИПСИНА, СПОСОБ ЕГО ПОЛУЧЕНИЯ И ПРИМЕНЕНИЯ

Изобретение относится к области биотехнологии, конкретно к получению варианта альфа-1-антитрипсина, и может быть использовано в медицине для предотвращения или лечения дефицита альфа-1-антитрипсина. Получают вариант альфа-1-антитрипсина путем замены аминокислоты в 4, 9 или 12 положении N-конца альфа-1-антитрипсина на аспарагин с целью добавления сайта гликозилирования. Изобретение позволяет увеличить период полужизни альфа-1-антитрипсина в организме. 7 н. и 1 з.п.ф-лы, 10 ил., 3 табл., 5 пр.

КОМПОЗИЦИИ ЛИПИДНЫХ НАНОЧАСТИЦ И СПОСОБЫ ДЛЯ ДОСТАВКИ мРНК

... 1. Композиция, включающая (а) участок, кодирующий функциональный секретируемый полипетпид; и (б) носитель-переносчик, включающий липидную наночастицу.2. Композиция по п.1, отличающаяся тем, что мРНК кодирует фермент, который находится в аномальном дефиците у индивидуума с лизосомной болезнью накопления.3. Композиция по п.1, отличающаяся тем, что мРНК кодирует функциональный эритропоэтин или функциональный полипептид α-галактозидазу.4. Композиция по п.1, отличающаяся тем, что молекула РНК включает по меньшей мере одну модификацию, которая придает молекуле РНК стабильность.5. Композиция по п.1, отличающаяся тем, что молекула РНК включает модификацию 5′ нетранслируемого участка указанной молекулы РНК.6. Композиция по п.5, отличающаяся тем, что указанная модификация включает введение структуры Cap1.7. Композиция по п.1, отличающаяся тем, что молекула РНК включает модификацию 3′ нетранслируемого участка указанной молекулы РНК.8. Композиция по п.7, отличающаяся тем, что указанная модификация ...

СПОСОБ ПОЛУЧЕНИЯ БЕЛКОВ В PICHIA PASTORIS, У КОТОРЫХ ОТСУТСТВУЕТ ПОДДАЮЩАЯСЯ ОБНАРУЖЕНИЮ ПЕРЕКРЕСТНАЯ СВЯЗУЮЩАЯ АКТИВНОСТЬ ПО ОТНОШЕНИЮ К АНТИТЕЛАМ ПРОТИВ АНТИГЕНОВ КЛЕТКИ-ХОЗЯИНА

... 1. Способ получения рекомбинантного гликопротеина в, у которого отсутствует поддающаяся обнаружению перекрестная связующая активность с антителами, полученными против антигенов клетки-хозяина, который включает:(a) предоставление рекомбинантной клетки-хозяина, которая не проявляет активность β-маннозилтрансферазы 2 в отношениигликана илигликана и не проявляет по меньшей мере одну активность, выбранную из активности β-маннозилтрансферазы 1 и активности β-маннозилтрансферазы 3 в отношениигликана илигликана, и которая содержит молекулу нуклеиновой кислоты, кодирующую рекомбинантный гликопротеин;(b) выращивание клетки-хозяина в среде в условиях, эффективных для экспрессии рекомбинантного гликопротеина; и(c) извлечение рекомбинантного гликопротеина из среды с получением рекомбинантного гликопротеина, у которого отсутствует поддающаяся обнаружению перекрестная связующая активность с антителами, полученными против антигенов клетки-хозяина.2. Способ по п.1, где клетка-хозяин не проявляет активность ...

СЛИТЫЕ СЕРПИНОВЫЕ ПОЛИПЕПТИДЫ И СПОСОБЫ ИХ ПРИМЕНЕНИЯ

СПОСОБ ПОЛУЧЕНИЯ РАСТВОРА АЛЬФА-1-АНТИТРИПСИНА

... 1. Способ очистки А1АТ из содержащих А1АТ растворов от других белковых компонентов, включающий стадии (a) проведения ионообменной хроматографии содержащего А1АТ раствора; (b) добавления детергентов и необязательно растворителя для инактивации покрытых липидной оболочкой вирусов; (c) последующего увеличения концентрации соли для высаливания детергентов. 2. Способ по п.1, в котором упомянутый содержащий А1АТ раствор получают из плазмы крови или ее фракций, предпочтительно из восстановленной плазматической фракции Коэна IV1, или получают из рекомбинантного или экспрессированного трансгенным путем препарата А1АТ или надосадочной жидкости после ферментации. 3. Способ по п.1 и/или 2, в котором ионообменную хроматографию выполняют на анионообменном геле, предпочтительно на DEAE-Sepharose® или DEAE-Sepharose® Fast Flow. 4. Способ по п.3, в котором упомянутую инактивацию вирусов в соответствии со стадией (b) выполняют путем добавления Triton X-100, Polysorbate 80 (Tween 80), TnBP и/или каприловой ...

VERFAHREN ZUR ABTRENNUNG DES ALPHA-1-PROTEINASE INHIBITORS VON EINER COHN-PASTE, WELCHE DIE FRAKTIONEN IV-1 + IV-4 ENTHÄLT

VERFAHREN ZUR ABTRENNEN VON ALPHA-1-PROTEINASE-INHIBITOR VON COHN-FRAKTION IV1-IV4 PASTE

VERFAHREN ZUR TRENNUNG VON ALPHA-1-PROTEINASE AUS BLUTPLASMAFRAKTIONEN.

Rheumatoid arthritis treatment

A synthetic peptide of not more than 20 amino acid residues or analogue thereof comprising a thiol-active cysteine residue and at least two positively charged amino acid residues situated on (but not necessarily adjacent to) the N-terminal side, on (but not necessarily adjacent to) the C-terminal side or on (but not necessarily adjacent to) both the N- and C- terminal sides of the thiol active cysteine for use in therapy.

RHEUMATOID ARTHRITIS TREATMENT

Methods for production of recombinant alpha1-antitrypsin

Methods of producing properly folded recombinant a l -antitrypsin (AAT) polypeptide are provided. Denatured recombinant AAT polypeptide is refolded by first solubilizing the polypeptide with a chaotroph at high pH, followed by refolding in the presence of reduced concentrations of chaotroph and in the presence of PEG, glycerol or sucrose, or a detergent while the pH is slowly reduced and is generally maintained.

MUTANT FUNGAL STRAIN DETECTION

Transgene expression

PROCEDURE FOR THE PREPARATION OF STABLE ZELLINIEN FROM TRANSGENEN ANIMALS FOR THE PRODUCTION OF CERTAIN PROTEINS, TUMORZELLINIEN AND FROM IT RECEIVED PROTEINS.

ALPHA 1-ANTITRYPSIN-PRÄPARATION SOWIE VERFAHREN ZU DEREN HERSTELLUNG

USE OF THE CRE-ERT2 OF FUSION PROTEIN FOR KONDITIONALEN PURPOSEFUL DNS RECOMBINATION IN THE MOUSE

ALPHA1-ANTITRYPSIN-ZUSAMMENSETZUNGEN AND TREATMENT PROCEDURE USING THESE COMPOSITIONS

SEPARATION FROM PLASMA COMPONENTS

ALPHA-1-ANTITRYPSINMUTANTE AND BUT CODING DNS AND PRODUCTION THE SAME, AND SUCH MUTANTE UTILIZATION THERAPEUTIC COMPOSITIONS AND YOUR PRODUCTION.

PROCEDURE FOR THE PRODUCTION OF A CONCENTRATE OF ALPHA-1-ANTITRYPSIN OF HUMAN PLASMA PARLIAMENTARY GROUP AND ITS USE AS DRUGS.

PROCEDURE FOR THE PRODUCTION OF HIGHLY PURIFIED ALPHA-1-PROTEINASE-INHIBITOR.

Alpha-1-antitrypsin compositions

A streamlined method for purifying alpha-1-antitrypsin (AAT) from an AAT-containing protein mixture, such as a Cohn fraction IV precipitate, is provided. In the method of the invention, contaminating proteins are destabilized by cleavage of disulfide bonds with a reducing reagent, such as a dithiol, which does not affect AAT. The destabilized proteins are then preferentially adsorbed on a solid protein-adsorbing material, without the addition of a salt as a precipitant. Separation of the solid adsorbent from the solution leaves a purified AAT solution that is directly suitable for chromatographic purification, without the need for extensive desalting as in prior art processes. A process incorporating this method, which provides pharmaceutical grade AAT in high yield on a commercial scale, is also described.

Adeno-associated virus serotype i nucleic acid sequences, vectors and host cells containing same

The nucleic acid sequences of adeno-associated virus (AAV) serotype 1 are provided, as are vectors and host cells containing these sequences and functional fragments thereof. Also provided are methods of delivering genes via AAV-1 derived vectors.

Polypeptides derived from alpha-1 antitrypsin and methods of use thereof

The present invention relates to isolated polypeptides comprising the amino acid sequence of residues 378-413 of Mus musculus α-1-antitrypsyn (serpina1c), and active fragments thereof, and to pharmaceutical compositions comprising same. The compositions of the invention are useful for treating burns, inflammatory, autoimmune and degenerative diseases.

Purification of cell culture derived alpha1 protease inhibitor

Described herein are methods for purifying recombinant, cell culture derived alpha 1 -protease inhibitor and removing a colored species that co-purifies with the recA1PI protein. Also described are methods for reducing the iron in cell culture derived alpha 1 -protease inhibitor.

COMPOSITIONS AND METHODS FOR TREATING PARKINSON'S DISEASE

Described herein are methods for treating a subject having or at risk of developing Parkinson's disease, by administering pluripotent cells that express glucocerebrosidase (GBA) or pluripotent cells that express GBA and one or more M2-promoting agents to the subject. Also disclosed are compositions comprising pluripotent cells expressing GBA, such as pluripotent cells expressing GBA and one or more M2-promoting agents. 1. A method of treating Parkinson's disease in a subject , the method comprising administering to the subject a composition comprising a population of pluripotent cells that express a transgene encoding glucocerebrosidase (GBA).2. The method of claim 1 , wherein the GBA is full-length GBA.3. The method of claim 1 , wherein the GBA is a catalytic domain of GBA.4. The method of any one of - claim 1 , wherein the GBA has an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO. 1.5. The method of claim 4 , wherein the GBA has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO. 1.6. The method of claim 5 , wherein the GBA has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO. 1.7. The method of claim 6 , wherein the GBA has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO. 1.8. The method of any one of - claim 6 , wherein the GBA has an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO. 5.9. The method of claim 8 , wherein the GBA has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO. 5.10. The method of claim 9 , wherein the GBA has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO. 5.11. The method of claim 10 , wherein the GBA has the amino acid sequence of SEQ ID NO. 5.12. The method of any one of - claim 10 , wherein the transgene encoding GBA has a nucleic ...

SERPIN FUSION POLYPEPTIDES AND METHODS OF USE THEREOF

This invention relates to molecules, particularly polypeptides, more particularly fusion proteins that include a serpin polypeptide or an amino acid sequence that is derived from a serpin and second polypeptide comprising of at least one the following: an Fc polypeptide or an amino acid sequence that is derived from an Fc polypeptide; a cytokine targeting polypeptide or a sequence derived from a cytokine targeting polypeptide; a WAP domain containing polypeptide or a sequence derived from a WAP containing polypeptide; and an albumin polypeptide or an amino acid sequence that is derived from a serum albumin polypeptide. This invention also relates to methods of using such molecules in a variety of therapeutic and diagnostic indications, as well as methods of producing such molecules. 1. A method of treating or alleviating a symptom of a disease or disorder associated with aberrant serine protease expression or activity in a subject in need thereof , the method comprising administering an isolated fusion protein comprising at least one alpha-1 antitrypsin (AAT) polypeptide , an Elafin polypeptide , and an immunoglobulin Fc polypeptide , such that at least two of the AAT polypeptide , the Elafin polypeptide , and the immunoglobulin Fc polypeptide are operably linked via a hinge region , a linker region , or both a hinge region and linker region.2. A method of treating or alleviating inflammation or a symptom of an inflammatory disease or disorder while reducing the risk of infection , in a subject in need thereof , the method comprising administering an isolated fusion protein comprising at least one alpha-1 antitrypsin (AAT) polypeptide , an Elafin polypeptide , and an immunoglobulin Fc polypeptide , such that at least two of the AAT polypeptide , the Elafin polypeptide , and the immunoglobulin Fc polypeptide are operably linked via a hinge region , a linker region , or both a hinge region and linker region.3. A method of reducing the risk of infection in a subject in ...

METHODS FOR EXTRACTING PROTEINS FROM BLOOD PLASMA

A method of producing protein products including alpha-1-proteinase inhibitor, gamma globulin, albumin, and other proteins from plasma includes steps of: (1) adding a salt to the blood product to produce a first intermediate, wherein the salt comprises between 11-13 wt % of the first intermediate; (2) separating the first intermediate to produce a first supernatant and a first paste; (3) adding a salt to the first intermediate to produce a second intermediate, wherein the salt comprises between 21-23 wt % of the second intermediate; (4) separating the second intermediate to produce a second supernatant and a second paste; (5) separating a third intermediate from the second supernatant by affinity chromatography; and (6) separating the third intermediate by ion exchange chromatography to produce an eluate containing the protein product. Advantageously, the inventive methods are simple and produce alpha-1-proteinase inhibitor, gamma globulin, albumin, and other proteins in high yields. 1. A method of separating A1P1 from a blood plasma containing product , the method comprising:thawing a frozen blood plasma product followed by stirring at a temperature suitable for dissolving a cryoprecipitate to generate the blood plasma containing product;adding a salt to the blood plasma containing product to produce a first intermediate, wherein the salt comprises between 11-20 wt % of the first intermediate;separating the first intermediate to produce a first supernatant and a first paste;adding a salt to the first supernatant to produce a second intermediate, wherein the salt comprises between 15-30 wt % of the second intermediate;separating the second intermediate to produce a second supernatant and a second paste;separating the second supernatant by affinity chromatography using an A1P1-specific affinity media to generate a flow-through fraction and a first eluate, wherein the first eluate comprises A1P1.2. The method of claim 1 , wherein the step of adding the salt to the ...

COMPOSITIONS, METHODS AND USES FOR ALPHA-1 ANTITRYPSIN OR DERIVATIVES THEREOF

Embodiments herein report methods and compositions for treating or preventing adverse effects of radiation therapies. In certain embodiments, compositions and methods relate to reducing or inhibiting damage due to acute, periodic or chronic radiation exposure. 1. A method for ameliorating adverse effects of radiation exposure in a subject comprising , administering to a subject undergoing or scheduled to undergo radiation therapy a therapeutically effective amount of a composition comprising alpha-1 antitrypsin (AAT) , a fragment thereof or a mutant thereof wherein the composition modulates the adverse effects of the radiation therapy in the subject.2. The method of claim 1 , wherein the composition comprises naturally occurring AAT (SEQ ID NO:1 or SEQ ID NO:39).3. The method of claim 1 , wherein the composition comprises a composition of one or more carboxyterminal fragments of naturally occurring AAT.4. The method of claim 1 , wherein the subject is undergoing radiation therapy for a cancer.5. The method of claim 1 , wherein the subject has been exposed to radiation as a result of a nuclear accident claim 1 , nuclear test claim 1 , nuclear attack claim 1 , or having undergone or is undergoing a diagnostic procedure comprising exposure to radiation.6. The method of claim 1 , wherein the composition is administered in a therapeutically effective amount to reduce or prevent at least one radiation-induced effect selected from the group consisting of myelosuppression claim 1 , renal toxicity claim 1 , weight loss claim 1 , behavioral changes claim 1 , pain claim 1 , nausea claim 1 , vomiting claim 1 , diarrhea claim 1 , constipation claim 1 , anemia claim 1 , malnutrition claim 1 , hair loss claim 1 , numbness claim 1 , changes in tastes claim 1 , loss of appetite claim 1 , thinned or brittle hair claim 1 , mouth sores claim 1 , memory loss claim 1 , hemorrhage claim 1 , cardiotoxicity claim 1 , hepatotoxicity claim 1 , ototoxicity claim 1 , and post-chemotherapy ...

Modified serpins for the treatment of bradykinin-mediated disease

The present invention relates to modified serpins for use in the treatment of bradykinin-mediated diseases. The modified serine protease inhibitors (serpins) have mutations in one or more of the P4, P3, P2, P1 and P1′ residues of their reactive center loop, which mutations increase the serpin's inhibition of plasma kallikrein (PK) as compared to the corresponding unmodified serpin. The mutations in the modified serpins of the invention further ensure that serpins display substantially no inhibition of at least thrombin and activated protein C. A modified serpin of the invention further preferably shows increased inhibition of at least one of an active form of Factor XII (FXII) and plasmin as compared to the corresponding unmodified serpin, and, preferably, the serpin inhibits at least one of an active form of FXII and PK stronger than they are inhibited by C1 esterase inhibitor. Preferably the modified serpin is a modified α1-antitrypsin. The invention further pertains to nucleic acid molecule encoding the modified serpins of the invention, e.g. a gene therapy vector, and to pharmaceutical compositions comprising the modified serpins of the invention or such gene therapy vectors. 115.-. (canceled)16. A method of treating a bradykinin-mediated disease , wherein the method comprises administering to a subject in need thereof an effective amount of a modified serpin wherein the serpin has mutations in one or more of the P4 , P3 , P2 , P1 and P1′ residues of its reactive center loop (RCL) , wherein the P1 residue is lysine or arginine , the P2 residue is not proline and wherein the P4 residue is serine , wherein the mutations increase the inhibition of plasma kallikrein (PK) as compared to the corresponding unmodified serpin , wherein the serpin more strongly inhibits PK than the serpin inhibits thrombin , wherein preferably , the serpin more strongly inhibits PK than the serpin inhibits activated protein C (APC) and wherein the serpin comprises an amino acid sequence ...

COMPOSITIONS AND METHODS FOR PREPARING A SUBJECT FOR ORGAN OR NON-ORGAN IMPLANTATION

Embodiments of the present invention illustrate methods of preventing transplantation rejection. In certain embodiments, a subject in need of an organ or non-organ transplantation can be pretreated with an AAT composition to reduce the incidence of transplantation rejection in the subject. Other embodiments include treating a subject with a composition including AAT before, during or after plastic surgery. 1. A method for reducing transplantation rejection in a subject , the method comprising administering to the subject at least 9 weeks prior to transplantation a composition comprising alpha1-antitrypsin (AAT) , a cleavage product thereof , a recombinant or fusion AAT molecule thereof; and a therapeutically acceptable excipient; and reducing transplantation rejection in the subject.2. The method of claim 1 , wherein the composition further comprises one or more anti-transplant rejection agent claim 1 , anti-inflammatory agent claim 1 , immunosuppressive agent claim 1 , immunomodulatory agent claim 1 , anti-microbial agent claim 1 , anti-viral agent or a combination thereof.3. The method of claim 1 , wherein the AAT comprises naturally-occurring full-length AAT and is administered at about 80 to about 150 mg/kg per dose.4. The method of claim 1 , wherein the AAT comprises an AAT fusion molecule and is administered at about 0.01 to about 10 mg/kg.5. The method of claim 1 , wherein the transplant is selected from an organ or non-organ transplant.6. The method of claim 5 , wherein the organ transplant is selected from the group consisting of lung claim 5 , kidney claim 5 , heart claim 5 , liver claim 5 , soft tissue claim 5 , skin claim 5 , pancreas claim 5 , intestine and a combination thereof.7. The method of claim 5 , wherein the non-organ transplant is selected from the group consisting of cornea claim 5 , skin grafting claim 5 , bone marrow claim 5 , stem claim 5 , pancreatic islet claim 5 , other transplanted cells and a combination thereof.8. The method of claim ...

PEPTIDES AND METHODS OF USING SAME

We describe peptides and their uses for the treatment of autoimmune, inflammatory and metabolic diseases. 1. A method of improving glycemic control in a subject having hyperglycemia comprising administering to the subject having hyperglycemia an oral formulation comprising a peptide selected from the group consisting of:(a) a peptide consisting of the amino acid sequenceX1-Z1-F-N-K-P-F-X2-Z2-X3-Z3-Q (SEQ ID NO: 2), whereinX1 is V or L;X2 is V, L or M;X3 is M, I or V;Z1 is any amino acid;Z2 is a sequence of any two amino acids; andZ3 is a sequence any five amino acids, and wherein the peptide is 37 or fewer amino acids;(b) a peptide consisting of the amino acid sequence VKFNKPFVFLMIEQNTK (SEQ ID NO: 1);(c) a peptide consisting essentially of the amino acid sequenceX1-Z1-F-N-X2-P-F-X3-Z2-X4-Z3-X5 (SEQ ID NO: 3), whereinX1 is V or L;X2 is K or R;X3 is V, L or M;X4 is M, I or V;X5 is K or Q;Z1 is any amino acid;Z2 is a sequence of any two amino acids; andZ3 is a sequence any five amino acids;(d) a peptide consisting essentially of the amino acid sequence RFNRPFLR (SEQ ID NO: 4).(e) a peptide consisting essentially of the amino acid sequence of RRRFNRPFLRRR (SEQ ID NO: 8).(f) a peptide consisting essentially of the amino acid sequence of VKFNKPFVFLMIEQNTK (SEQ ID NO: 1); and(g) a peptide consisting essentially of the amino acid sequence of FNRPFL (SEQ ID NO: 10).2. The method of claim 1 , wherein the peptide further comprises at least one second peptide or protein.3. The method of claim 2 , wherein the at least one second protein or peptide is attached to the peptide as a fusion peptide.4. The method of claim 2 , wherein the at least one second peptide or protein is an epitope tag or a half-life extender or both.5. The method of claim 1 , wherein the peptide comprises one or more D-amino acids.6. The method of claim 1 , wherein the subject having hyperglycemia is affected with type II diabetes or type I diabetes.7. The method of claim 6 , wherein the inflammatory ...

Lipid nanoparticle compositions and methods for mrna delivery

Disclosed herein are compositions and methods for modulating the production of a protein in a target cell. The compositions and methods disclosed herein are capable of ameliorating diseases associated with protein or enzyme deficiencies.

SERPIN FUSION POLYPEPTIDES AND METHODS OF USE THEREOF

This invention relates to molecules, particularly polypeptides, more particularly fusion proteins that include a serpin polypeptide or an amino acid sequence that is derived from a serpin and second polypeptide comprising of at least one the following: an Fc polypeptide or an amino acid sequence that is derived from an Fc polypeptide; a cytokine targeting polypeptide or a sequence derived from a cytokine targeting polypeptide; a whey acidic protein (WAP) domain containing polypeptide or a sequence derived from a WAP containing polypeptide; and an albumin polypeptide or an amino acid sequence that is derived from a serum albumin polypeptide. This invention also relates to methods of using such molecules in a variety of therapeutic and diagnostic indications, as well as methods of producing such molecules. 1. An isolated fusion protein comprising at least one human serpin polypeptide operably linked to a second polypeptide , wherein the second polypeptide comprises an albumin polypeptide or an amino acid sequence that is derived from a serum albumin polypeptide.2. An isolated fusion protein comprising at least one alpha-1 antitrypsin (AAT) polypeptide operably linked to a second polypeptide , wherein the second polypeptide comprises a human serum albumin (HSA) polypeptide.3. The fusion protein of claim 1 , wherein the human serpin polypeptide is a human alpha-1 antitrypsin (AAT) polypeptide or is derived from a human AAT polypeptide.4. The isolated fusion protein of any one of - claim 1 , wherein AAT polypeptide comprises the amino acid sequence of SEQ ID NO: 2.5. The isolated fusion protein of any one of - claim 1 , wherein the AAT polypeptide comprises the reactive site loop of AAT comprising the amino acid sequence of SEQ ID NO: 1 or a mutated reactive site loop of AAT comprising the amino acid sequence of SEQ ID NO: 32 or 33.6. The fusion protein of claim 1 , wherein the albumin polypeptide is a human serum albumin (HSA) polypeptide or is derived from a HSA ...

Methods for single chain variable region enox2 antibodies for cancer detection and diagnosis

Cancers of different cellular or tissue origins express different ENOX2 cancer isoforms or combinations of isoforms and shed these proteins into the circulation. Herein are disclosed methods both for cancer detection and diagnosis of particular origin, based on the patterns and molecular weights of the isoforms which allow the identification of the cell type and or tissue of origin of the neoplasm. Relative ENOX2 amounts are proportional to tumor burden and provide a reliable measure of response to therapy and disease progression. Also provided is the amino acid sequence to which the scFv antibodies bind as the molecular basis for the specificity of the test.

Recombinant Glycoproteins with Reduced Antennary Fucosylation

The present invention relates to methods for reducing antennary fucosylation of complex N-glycans in recombinantly expressed glycoproteins, cell lines that can be used in said methods, respective recombinant glycoproteins, and methods for expressing the same in said cell lines.

METHODS AND COMPOSITIONS FOR ALPHA-1 ANTITRYPSIN RELATED DISEASE DISORDERS

The invention relates to methods and compositions directed at obtaining a non-natively glycosylated recombinant human alpha-1 antitrypsin (A1AT) peptides that are glycosylated in a non-native configuration that confer enhanced biologic activities. 1. A recombinant glycoprotein comprising human alpha-1 antitrypsin having non-native glycosylation.2. The recombinant glycoprotein according to claim 1 , wherein said glycoprotein comprises SEQ ID NO: 18 claim 1 , SEQ ID NO: 19 claim 1 , or SEQ ID NO: 20.3. The recombinant glycoprotein according to claim 1 , wherein said glycosylation comprises a population of substantially homogeneous N-glycans.4. (canceled)5. The recombinant glycoprotein according to claim 1 , wherein said glycosylation comprises one or more of Man5GlcNAc2 claim 1 , Man6GlcNAc2 claim 1 , Man7GlcNAc2 claim 1 , Man8GlcNAc2 claim 1 , Man9GlcNAc2 claim 1 , GlcNAcMan5GlcNAc2 claim 1 , GalGlcNAcMan5GlcNAc2 claim 1 , GalGlcNAcMan3GlcNAc2 claim 1 , GlcNAcMan3GlcNAc2 claim 1 , GlcNAc2Man3GlcNAc2 claim 1 , Gal2GlcNAc2Man3GlcNAc2 claim 1 , Hex10GlcNac2 claim 1 , Hex11GlcNac2 claim 1 , Hex12GlcNac2 claim 1 , Hex13GlcNac2 claim 1 , Hex14GlcNac2 claim 1 , Hex15GlcNac2 claim 1 , and Hex16GlcNac2.68-. (canceled)9. The recombinant glycoprotein according to claim 1 , wherein Man7GlcNAc2 claim 1 , Man8GlcNAc2 claim 1 , and Man9GlcNAc2 N-glycans comprise less than 20% claim 1 , less than about 10% claim 1 , less than about 5% claim 1 , or less than about 1% of the total N-glycans.10. The recombinant glycoprotein according to claim 1 , wherein Hex10GlcNac2 claim 1 , Hex11GlcNac2 claim 1 , Hex12GlcNac2 claim 1 , Hex13GlcNac2 claim 1 , Hex15GlcNac2 claim 1 , and Hex16GlcNac2 N-glycans comprise less than about 20% claim 1 , less than about 10% claim 1 , less than about 5% claim 1 , or less than about 1% of the total N-glycans.11. The recombinant glycoprotein according to claim 1 , wherein HexGlcNac2 N-glycans comprise less than about 20% claim 1 , less than about 10% claim 1 , ...

COMPOSITIONS AND METHODS FOR THE DIAGNOSIS OF RHEUMATOID ARTHRITIS

The present disclosure relates to the field of molecular biology and more specifically to methods for detecting anti-carbamylated protein (anti-CarP) antibodies in the serum of rheumatoid arthritis (RA) patients. 1. A purified polypeptide comprising an in vitro carbamylated human alpha 1 antitrypsin (hA1AT) , or an in vitro carbamylated fragment thereof , wherein the purified polypeptide is immobilized on a solid support.2. The purified polypeptide of claim 1 , wherein the purified polypeptide is a purified recombinant polypeptide encoded by cDNA claim 1 , or the purified polypeptide is hA1AT claim 1 , or an in vitro carbamylated fragment thereof claim 1 , purified from blood claim 1 , serum claim 1 , plasma claim 1 , urine claim 1 , or synovial fluid claim 1 , or wherein the hA1AT claim 1 , or the in vitro carbamylated fragment thereof claim 1 , comprises the amino acid sequence of SEQ ID NO:1 claim 1 , or wherein the hA1AT claim 1 , or the in vitro carbamylated fragment thereof claim 1 , has greater than 70% claim 1 , greater than 75% claim 1 , greater than 80% claim 1 , greater than 85% claim 1 , greater than 90% claim 1 , greater than 95% claim 1 , greater than 96% claim 1 , greater than 97% claim 1 , greater than 98% claim 1 , or greater than 99% sequence identity to SEQ ID NO:1 claim 1 , or wherein the hA1AT claim 1 , or the in vitro carbamylated fragment thereof claim 1 , comprises a fragment of 8 or more contiguous amino acids of SEQ ID NO:1 claim 1 , or wherein the hA1AT claim 1 , or the in vitro carbamylated fragment thereof claim 1 , comprises a fragment of 8 or more contiguous amino acids with greater than 80% claim 1 , greater than 85% claim 1 , greater than 90% claim 1 , greater than 95% claim 1 , greater than 96% claim 1 , greater than 97% claim 1 , greater than 98% claim 1 , or greater than 99% sequence identity to SEQ ID NO:1 claim 1 , or wherein the hA1AT claim 1 , or the in vitro carbamylated fragment thereof claim 1 , comprises the amino acid ...

SERUM-FREE STABLE TRANSFECTION AND PRODUCTION OF RECOMBINANT HUMAN PROTEINS IN HUMAN CELL LINES

The present invention relates to an improved method for the serum-free production of an immortalized human cell line stably transfected under serum-free conditions with a specific vector carrying the gene coding for the protein of interest. Furthermore the invention relates to a production cell line obtained by said method, a production method for said protein of interest utilizing said production cell line, and the specific vector carrying the gene of interest itself. 1. A stably transfected immortalized human cell line prepared by transfecting an immortalized human host cell line under serum-free conditions with a transfection vector comprisinga nucleic acid sequence comprising a gene encoding a human target protein or a derivative or mutant thereof, a promoter, and a bovine growth hormone polyadenylation (polyA) signal, the promoter and polyA signal linked to the 5′ and 3′ end of the gene encoding the human target protein, respectively, andan origin of replication,resulting in a stably transfected immortalized human cell line.2. The stably transfected immortalized human cell line of where the human cell line is 293F and the transfection vector is derived from pcDNA3.1.3. The stably transfected immortalized human cell line of claim 1 , where the human target protein is a human plasma protein selected from the group consisting ofblood clotting factors selected from the group consisting of factor IX, factor VIII (wild-type and B-domain deleted), Factor VII/VIIa, and von Willebrand factor (vWF);growth factors including erythropoietin;colony-stimulating factors (CSFs) selected from the group consisting of granulocyte stimulating factor (G-CSF), macrophage CSF (M-CSF), and granulocyte-macrophage CSF (GM-CSF);cytokines including interleukins;protease inhibitors selected from the group consisting of alpha-1-antitrypsin (A1AT) and chymotrypsin;transport proteins selected from the group consisting of hormones, inhibitory or regulatory acting proteins, and derivatives and ...

METHODS OF TREATMENT USING ALPHA-1-ANTITRYPSIN COMPOSITIONS

A streamlined method for purifying alpha-1-antitrypsin (AAT) from an AAT-containing protein mixture, such as Coh fraction IV precipitate, is provided. In the method of the invention, contaminating proteins are destabilized by cleavage of disulfide bonds with a reducing reagent, such as dithiol, which does not affect AAT. The destabilized proteins are then preferentially adsorbed on a solid protein-adsorbing material, without the addition of a salt as a precipitante. Spearation of the solid absorbent from the solution leaves a purified AAT solution that is directly suitable for chromatographic purification, without the need for extensive desalting as in prior art processes. A process incorporating this method, which provides pharmaceutical grade AAT in high yield on a commercial scale, is also described. 130-. (canceled)31. A method of treating alpha-1-antitrypsin (AAT) deficiency in a subject in need thereof , comprising administering to the subject a pharmaceutically effective amount of a pharmaceutical composition comprising plasma-derived alpha-1-antitrypsin (AAT) , wherein said composition further comprises:(a) less than 0.1% albumin;{'sub': '1', '(b) less than 0.8% α-acid glycoprotein;'}{'sub': '2', '(c) less than 0.1% α-macroglobulin;'}(d) less than 0.1% apolipoprotein A1;(e) less than 0.5% antithrombin III; and(f) less than 0.1% ceruloplasmin.32. The method of claim 31 , wherein the apparent ratio of active to antigenic AAT is greater than 1.08 when measured by endpoint nephelometry.33. The method of claim 31 , wherein the specific activity of the AAT is at least 0.99 mg functional AAT per milligram of protein claim 31 , when using as an extinction coefficient E=5.3.34. The method of claim 31 , wherein the specific activity of the AAT is less than or equal to 1.12 mg functional alpha-1-antitrypsin per milligram of protein.35. The method of claim 31 , wherein enveloped viruses are reduced in number by at least 11 logunits and non-enveloped viruses are reduced ...

MESSENGER UNA MOLECULES AND USES THEREOF

This invention provides a range of translatable messenger UNA (mUNA) molecules. The mUNA molecules can be translated in vitro and in vivo to provide an active polypeptide or protein, or to provide an immunization agent or vaccine component. The mUNA molecules can be used as an active agent to express an active polypeptide or protein in cells or subjects. Among other things, the mUNA molecules are useful in methods for treating rare diseases. 1. A mUNA molecule , comprising one or more UNA monomers , and comprising nucleic acid monomers , wherein the mUNA molecule is translatable to express a polypeptide or protein.2. The molecule of claim 1 , wherein the molecule comprises from 200 to 12 claim 1 ,000 monomers.3. The molecule of claim 1 , wherein the molecule comprises from 200 to 4 claim 1 ,000 monomers.4. The molecule of claim 1 , wherein the molecule comprises from 1 to 8 claim 1 ,000 UNA monomers.5. The molecule of claim 1 , wherein the molecule comprises from 1 to 100 UNA monomers.6. The molecule of claim 1 , wherein the molecule comprises from 1 to 20 UNA monomers.7. The molecule of claim 1 , wherein the molecule comprises one or more modified nucleic acid nucleotides claim 1 , or one or more chemically-modified nucleic acid nucleotides.8. The molecule of claim 1 , wherein the molecule comprises a 5′ cap claim 1 , a 5′ untranslated region of monomers claim 1 , a coding region of monomers claim 1 , a 3′ untranslated region of monomers claim 1 , and a tail region of monomers.9. The molecule of claim 8 , wherein the molecule comprises a translation enhancer in a 5′ or 3′ untranslated region.10. The molecule of claim 1 , wherein the molecule is translatable in vivo.11. The molecule of claim 1 , wherein the molecule is translatable in vitro.12. The molecule of claim 1 , wherein the molecule is translatable in a mammalian cell.13. The molecule of claim 1 , wherein the molecule is translatable in a human in vivo.14. The molecule of claim 1 , wherein a translation ...

METHODS AND COMPOSITIONS FOR MODULATING ALPHA-1-ANTITRYPSIN EXPRESSION

Disclosed herein are methods for decreasing A1AT mRNA and protein expression and treating, ameliorating, preventing, slowing progression, or stopping progression of fibrosis. Disclosed herein are methods for decreasing A1AT mRNA and protein expression and treating, ameliorating, preventing, slowing progression, or stopping progression of liver disease, such as, A1ATD associated liver disease, and pulmonary disease, such as, A1ATD associated pulmonary disease in an individual in need thereof. Methods for inhibiting A1AT mRNA and protein expression can also be used as a prophylactic treatment to prevent individuals at risk for developing a liver disease, such as, A1ATD associated liver disease and pulmonary disease, such as, A1ATD associated pulmonary disease. 1. A compound comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and comprising a nucleobase sequence comprising a portion of at least 8 , contiguous nucleobases complementary to an equal length portion of nucleobases 459 to 513 , 1349 to 1597 , 1561 to 1597 , 1564 to 1583 , 1575 to 1594 of SEQ ID NO: 1 , wherein the nucleobase sequence of the modified oligonucleotide is at least 90% complementary to SEQ ID NO: 1.29-. (canceled)10. The compound of claim 1 , wherein the nucleobase sequence of the modified oligonucleotide is at least 95% complementary to SEQ ID NO: 1.1135-. (canceled)36. A method of reducing A1AT in an animal comprising administering to the animal a modified oligonucleotide targeting an A1AT nucleic acid sequence as shown in SEQ ID NO: 1.37. The method of claim 36 , wherein the modified oligonucleotide targeting A1AT consists of 12 to 30 linked nucleosides and is at least 90% complementary to the A1AT nucleic acid.38. A method of treating claim 36 , ameliorating and/or preventing an A1ATD associated liver disease in an animal at risk for the A1ATD associated liver disease comprising claim 36 ,(a) identifying the animal at risk for developing the A1ATD associated liver ...

LIPID NANOPARTICLE COMPOSITIONS AND METHODS FOR MRNA DELIVERY

Disclosed herein are compositions and methods for modulating the production of a protein in a target cell. The compositions and methods disclosed herein are capable of ameliorating diseases associated with protein or enzyme deficiencies. 140-. (canceled)41. A method for delivery of messenger RNA (mRNA) for in vivo production of a IL-12 polypeptide , comprising administering , to a human , a composition comprising an mRNA that encodes the IL-12 polypeptide wherein the mRNA is encapsulated within a lipid nanoparticle , wherein the administering of the composition results in expression of the IL-12 polypeptide encoded by the mRNA that is detectable in a target tissue or in serum at least 72 hours after administration , and wherein the lipid nanoparticle comprises one or more PEG-modified lipids.42. The method of claim 41 , wherein the mRNA comprises a 5′ untranslated region.43. The method of claim 41 , wherein the mRNA comprises a 3′ untranslated region.44. The method of claim 43 , wherein the mRNA comprises a cap structure.45. The method of claim 44 , wherein the mRNA comprises a poly A tail.46. The method of claim 45 , wherein the lipid nanoparticle comprises one or more cationic lipids.47. The method of claim 46 , wherein the lipid nanoparticle comprises one or more non-cationic lipids.48. The method of claim 47 , wherein the mRNA is unmodified.49. The method of claim 48 , wherein the composition is a reconstituted lyophilized composition.50. The method of claim 47 , wherein the modification comprises a modified nucleotide.51. The method of claim 50 , wherein the modified nucleotide is pseudouridine.52. The method of claim 51 , wherein the composition is a reconstituted lyophilized composition.53. The method of claim 48 , wherein the composition is administered by intravenous injection.54. The method of claim 51 , wherein the composition is administered by intravenous injection.55. The method of claim 48 , wherein the composition is administered by intramuscular ...

MODULATION OF LIVER GENES

Described herein are compositions and methods for modulation of gene expression in the liver including modulation of PCSK9, TTR, SERPINA1, KLKB1 and/or HAO1. 2. The liver cell of claim 1 , wherein the sequence of the endogenous gene is altered by cleaving the gene using a pair of zinc finger nucleases (ZFNs) claim 1 , wherein at least one of the zinc finger nucleases of the pair comprises the zinc finger protein—and a cleavage domain.3. The liver cell of claim 2 , wherein the pair of ZFNs bind to SEQ ID NO:65 and SEQ ID NO:66; SEQ ID NO:220 and SEQ ID NO:221; SEQ ID NO:86 and SEQ ID NO:87 or SEQ ID NO:222; SEQ ID NO:232 and SEQ ID NO:233; SEQ ID NO:239 and SEQ ID NO:240; SEQ ID NO:241 and SEQ ID NO:242; SEQ ID NO:243 and SEQ ID NO:244; SEQ ID NO:245 and SEQ ID NO:246; or SEQ ID NO:247 and SEQ ID NO:248.4. The liver cell of claim 2 , further comprising integrating an exogenous sequence into the cleaved endogenous gene.5. The liver cell of claim 4 , wherein the exogenous sequence comprises a transgene; introduces a mutation into the gene claim 4 , or corrects a mutation in the endogenous gene.6. The liver cell of claim 1 , wherein the liver cell further comprises an artificial transcription factor comprising a DNA-binding domain and a transcriptional regulatory domain claim 1 , wherein the artificial transcription factor alters expression of the endogenous gene.7. The liver cell of claim 6 , wherein the artificial transcription factor activates or represses expression of the endogenous gene.8. A pharmaceutical composition comprising the liver cell according to .9. The pharmaceutical composition of claim 8 , further comprising genetically modified cells descended from the liver cell.10. A method of producing the liver cell according to claim 1 , the method comprising administering an artificial transcription factor or artificial nuclease that alters expression of the endogenous gene to the liver cell.12. The fusion molecule of claim 11 , wherein the functional domain ...

COMPOSITIONS CONTAINING ALPHA-1-ANTITRYPSIN AND METHODS FOR USE

Methods and compositions for treating patients (e.g., patients who are insulin resistant, patients who have diabetes, or are at risk for developing diabetes) are disclosed herein. The methods can include administration of an al antitrypsin (AAT) polypeptide or an agent, such as a nucleic acid molecule or organic compound, that promotes the expression or activity of α1-antitrypsin. 170-. (canceled)71. A method of transplanting a cell into a subject , the method comprising:transplanting a cell into the subject; andadministering to the subject a therapeutically effective amount of an α1-antitrypsin polypeptide or a nucleic acid molecule that encodes the α1-antitrypsin polypeptide, wherein the α1-antitrypsin polypeptide has at least 95% identity to a sequence of a wild type α1-antitrypsin polypeptide.72. The method of claim 71 , wherein the cell is an islet cell.73. The method of claim 71 , wherein the α1-antitrypsin polypeptide or a nucleic acid molecule that encodes the α1-antitrypsin polypeptide is administered topically claim 71 , intraperitoneally claim 71 , intravenously claim 71 , subcutaneously claim 71 , intrathecally claim 71 , intramuscularly claim 71 , intranasally claim 71 , orally claim 71 , transepidermally claim 71 , parenterally claim 71 , intracerebroventricularly claim 71 , or by inhalation.74. The method of claim 71 , wherein the α1-antitrypsin polypeptide is a full-length α1-antitrypsin polypeptide.75. The method of claim 74 , wherein the full-length α1-antitrypsin polypeptide is a human α1-antitrypsin polypeptide.76. The method of claim 71 , wherein the subject is administered a therapeutically effective amount of an α1-antitrypsin polypeptide that has at least 99% identity to a sequence of a wild type α1-antitrypsin polypeptide.77. The method of claim 71 , wherein the α1-antitrypsin polypeptide is joined to a portion of an immunoglobulin or an albumin.78. The method of claim 77 , wherein the portion of the immunoglobulin is an Fc region of an IgG ...

COMPOSITIONS CONTAINING ALPHA-1-ANTITRYPSIN AND METHODS FOR USE

Methods and compositions for treating patients (e.g., patients who are insulin resistant, patients who have diabetes, or are at risk for developing diabetes) are disclosed herein. The methods can include administration of an α1 antitrypsin (AAT) polypeptide or an agent, such as a nucleic acid molecule or organic compound, that promotes the expression or activity of α1-antitrypsin. 170-. (canceled)71. A nucleic acid molecule encoding a recombinant polypeptide comprising an α1-antitrypsin polypeptide conjugated to an Fc region of an immunoglobulin.72. A recombinant polypeptide comprising an α1-antitrypsin polypeptide conjugated to an Fc region of an immunoglobulin.73. A pharmaceutical composition comprising:{'claim-ref': {'@idref': 'CLM-00072', 'claim 72'}, 'the recombinant polypeptide of ; and'}a pharmaceutically acceptable carrier.74. A method of treating Type 1 diabetes in a subject , the method comprising:(a) identifying a subject in need of treatment for Type 1 diabetes; and(b) administering to the subject a therapeutically effective amount of an α1-antitrypsin polypeptide or an agent that promotes the expression or activity of α1-antitrypsin.75. The method of claim 74 , wherein the α1-antitrypsin polypeptide is a full-length α1-antitrypsin polypeptide or a biologically active fragment or mutant thereof.76. The method of claim 75 , wherein the full-length α1-antitrypsin polypeptide is a human α1-antitrypsin polypeptide.77. The method of claim 75 , wherein the full-length α1-antitrypsin polypeptide is conjugated to an Fc region of an immunoglobulin.78. The method of claim 75 , wherein the full-length α1-antitrypsin polypeptide is conjugated to an albumin.79. The method of claim 74 , wherein the agent that promotes the expression of α1-antitrypsin is a nucleic acid molecule encoding a full-length α1-antitrypsin polypeptide or a biologically active fragment or mutant thereof.80. The method of claim 74 , wherein the agent that promotes the activity of α1-antitrypsin ...

METHODS OF TREATMENT USING ALPHA-1-ANTITRYPSIN COMPOSITIONS

A streamlined method for purifying alpha-1-antitrypsin (AAT) from an AAT-containing protein mixture, such as Coh fraction IV precipitate, is provided. In the method of the invention, contaminating proteins are destabilized by cleavage of disulfide bonds with a reducing reagent, such as dithiol, which does not affect AAT. The destabilized proteins are then preferentially adsorbed on a solid protein-adsorbing material, without the addition of a salt as a precipitante. Spearation of the solid absorbent from the solution leaves a purified AAT solution that is directly suitabale for chromatographic purification, without the need for extensive desalting as in prior art processes. A process incorporating this method, which provides pharmaceutical grade AAT in high yield on a commercial scale, is also described. 130-. (canceled)31. A method of treating alpha-1-antitrypsin (AAT) deficiency in a subject in need thereof , comprising administering to the subject a pharmaceutically effective amount of a pharmaceutical composition comprising plasma-derived alpha-1-antitrypsin (AAT) , wherein the composition comprises:(a) 7.440 mg per unit dose of alpha-1-antitrypsin;(b) 0.059 mg sodium citrate; and(c) 0.001 mg citric acid; andwherein the composition is an aerosol.32. The method of claim 31 , wherein the composition is a dry powder.33. The method of claim 31 , wherein the composition is administered with a metered dose inhaler.34. The method of claim 31 , wherein the composition is administered with a pulmonary delivery device.35. The method of claim 31 , wherein the composition comprises 6 mg per unit dose of functional AAT.36. The method of claim 31 , wherein a delivered dose of 3.6 mg functional AAT is administered.37. The method of claim 31 , wherein the composition further comprises:(d) less than 0.1% albumin based on total protein weight; and(e) less than 0.1% apolipoprotein A1 based on total protein weight,{'sub': '1 cm,280 nm', 'sup': '1%', 'wherein total protein weight is ...

COMPOSITIONS OF, AND METHODS FOR, ALPHA-1 ANTI TRYPSIN Fc FUSION MOLECULES

A novel method of treating and preventing bacterial diseases is provided. In particular, the present invention relates to compositions and methods for inhibition of Gram negative, Gram positive and acid fast bacilli in general and tuberculosis (TB), complex (MAC), and anthrax in particular. Thus, the invention relates to modulation of cellular activities, including macrophage activity, and the like. More particularly, the present invention relates to the inhibitory compounds comprising naturally occurring and man-made inhibitors of serine protease. 1. A construct comprising a nucleic acid sequence encoding a fusion polypeptide comprising:a first nucleic acid sequence encoding mammalian alpha-1 antitrypsin (AAT), wherein the first nucleic acid sequence comprises SEQ ID NO: 68 or SEQ ID NO:78; anda second nucleic acid sequence encoding an IgG1 Fc.210-. (canceled)11. A method for treating a subject in need of c-1 antitrypsin (AAT) therapy , the method comprising administering to the subject a therapeutically effective amount of a composition comprising an AAT fusion polypeptide in an amount effective to treat the subject.12. The method according to claim 11 , wherein the AAT fusion polypeptide comprises a fusion molecule comprising a first polypeptide comprising mammalian alpha-1 antitrypsin (AAT) claim 11 , wherein the first polypeptide comprising mammalian AAT is represented by SEQ ID NO:69 or SEQ ID NO:79; and a second polypeptide comprising an IgG1 Fc claim 11 , an immunoglobulin constant region; and a pharmaceutically acceptable excipient thereof.13. The method according to claim 11 , wherein the second polypeptide comprising an IgG1 Fc is represented by SEQ ID NO:65.14. The method according to claim 11 , wherein the AAT fusion polypeptide comprises a fusion molecule comprising a first polypeptide comprising mammalian alpha-1 antitrypsin (AAT); and a second polypeptide comprising an IgG1 Fc claim 11 , an immunoglobulin constant region claim 11 , wherein the second ...

PROTEIN EXPRESSION ENHANCING POLYPEPTIDES

Fusion proteins comprising a protein expression enhancing polypeptide linked to a target protein binding domain and nucleic acid molecules encoding such fusion proteins are described for use in enhancing expression and/or location of a targeted protein of interest, for restoring lost functions in cells, and for treating disease. Additional fusion proteins comprising a target protein of interest modified with a fusion partner comprising a protein expression enhancing polypeptide are also disclosed. 1. A fusion protein comprising:(1) an isolated protein expression enhancing polypeptide (PEEP) domain linked to (2) a target protein of interest,wherein said isolated protein expression enhancing polypeptide domain (1) is selected from the group consisting of:(a) an isolated J domain of a J protein comprising a polypeptide domain of a J protein comprising four α helices, helices I, II, III, and IV, and having a histidine, proline, and aspartic acid sequence motif (HPD motif) between helix II and helix III, or an active fragment of said J domain; and X1 is isoleucine (I), leucine (L), valine (V), alanine (A), or methionine (M);', 'X2 and X3 are each independently any amino acid with the proviso that one or both are K or R;', 'X4 is any amino acid or X4 may be absent when X1 through X3 are present and X5 through X9 are present;', 'X5 is tyrosine (Y), tryptophan (W), or phenylalanine (F);', 'X6 and X7 are each independently any amino acid with the proviso that one or both are lysine (K) or arginine (R); or either one of X6 and X7 may be absent when the other is K or R and when X1 through X5 are present and X8 and X9 are present; and', 'X8 and X9 are any amino acid with the proviso that one or both are leucine (L) or alanine (A); or one of X8 and X9 may be absent when the other is L or A and when X1 through X7 are present; and, '(I) X1-X2-X3-X4-X5-X6-X7-X8-X9 (SEQ ID NO:47), wherein, '(b) a J domain analog polypeptide comprises the amino acid sequence of formula Iwherein said ...

COMPOSITIONS AND METHODS FOR THE DIAGNOSIS OF RHEUMATOID ARTHRITIS

The present disclosure relates to the field of molecular biology and more specifically to methods for detecting anti-carbamylated protein (anti-CarP) antibodies in the serum of rheumatoid arthritis (RA) patients. 1. A purified polypeptide comprising an in vitro carbamylated human alpha 1 antitrypsin (hA1AT) , or a fragment thereof.2. The purified polypeptide of claim 1 , wherein the purified polypeptide is a purified recombinant polypeptide encoded by cDNA.3. The purified polypeptide of claim 1 , wherein the purified polypeptide is hA1AT claim 1 , or a fragment thereof claim 1 , purified from blood claim 1 , serum claim 1 , plasma claim 1 , urine claim 1 , or synovial fluid.4. The purified polypeptide of claim 1 , wherein the hA1AT claim 1 , or fragment thereof claim 1 , comprises the amino acid sequence of SEQ ID NO:1.5. The purified polypeptide of claim 1 , wherein the hA1AT claim 1 , or fragment thereof claim 1 , has greater than 70% claim 1 , greater than 75% claim 1 , greater than 80% claim 1 , greater than 85% claim 1 , greater than 90% claim 1 , greater than 95% claim 1 , greater than 96% claim 1 , greater than 97% claim 1 , greater than 98% claim 1 , or greater than 99% sequence identity to SEQ ID NO:1.6. The purified polypeptide of claim 1 , wherein the hA1AT claim 1 , or fragment thereof claim 1 , comprises a fragment of 8 or more contiguous amino acids of SEQ ID NO:1.7. The purified polypeptide of claim 1 , wherein the hA1AT claim 1 , or fragment thereof claim 1 , comprises a fragment of 8 or more contiguous amino acids with greater than 80% claim 1 , greater than 85% claim 1 , greater than 90% claim 1 , greater than 95% claim 1 , greater than 96% claim 1 , greater than 97% claim 1 , greater than 98% claim 1 , or greater than 99% sequence identity to SEQ ID NO:1.8. The purified polypeptide of claim 1 , wherein the hA1AT claim 1 , or fragment thereof claim 1 , comprises the amino acid sequence of any one of SEQ ID NOs:3-32.9. The purified polypeptide of ...

LIPID NANOPARTICLE COMPOSITIONS AND METHODS FOR MRNA DELIVERY

Disclosed herein are compositions and methods for modulating the production of a protein in a target cell. The compositions and methods disclosed herein are capable of ameliorating diseases associated with protein or enzyme deficiencies. 140-. (canceled)41. A method of treating lung disease , comprising administering to a subject in need of treatment an effective amount of a composition comprising an mRNA encoding a protein or peptide , encapsulated within a lipid nanoparticle ,wherein the mRNA is encapsulated within a lipid nanoparticle comprising one or more PEG-modified lipids,wherein the composition is administered via pulmonary delivery, andwherein the administration of the composition results in detectable levels of the protein or peptide at least 72 hours after a single administration in the subject's lung.42. The method of claim 41 , wherein the composition comprises one or more cationic lipids.43. The method of claim 42 , wherein the one or more cationic lipids constitute from 2% to 70% of the total lipids by molar ratio.44. The method of claim 43 , wherein the one or more cationic lipids constitute from 5% to 50% of the total lipids by molar ratio.45. The method of claim 42 , wherein the one or more PEG-modified lipids constitute from 0.5% to 20% of the total lipids by a molar ratio.46. The method of claim 45 , wherein the one or more PEG-modified lipids constitute from 4% to 10% of the total lipids by molar ratio.47. The method of claim 45 , wherein the one or more cationic lipids comprise C12-200.48. The method of claim 45 , wherein the one or more cationic lipids comprise ICE.49. The method of claim 45 , wherein the one or more cationic lipids comprise HGT4003.50. The method of claim 47 , wherein the composition comprises one or more non-cationic lipids.51. The method of claim 48 , wherein the composition comprises one or more non-cationic lipids.52. The method of claim 51 , wherein the one or more non-cationic lipids constitute from 5% to 90% of the ...

Serpin fusion polypeptides and methods of use thereof

This invention relates to molecules, particularly polypeptides, more particularly fusion proteins that include a serpin polypeptide or an amino acid sequence that is derived from a serpin and second polypeptide comprising of at least one the following: an Fc polypeptide or an amino acid sequence that is derived from an Fc polypeptide; a cytokine targeting polypeptide or a sequence derived from a cytokine targeting polypeptide; a WAP domain containing polypeptide or a sequence derived from a WAP containing polypeptide; and an albumin polypeptide or an amino acid sequence that is derived from a serum albumin polypeptide. This invention also relates to methods of using such molecules in a variety of therapeutic and diagnostic indications, as well as methods of producing such molecules.

SERPIN FUSION POLYPEPTIDES AND METHODS OF USE THEREOF

This invention relates to molecules, particularly polypeptides, more particularly fusion proteins that include a serpin polypeptide or an amino acid sequence that is derived from a serpin and second polypeptide comprising of at least one the following: an Fc polypeptide or an amino acid sequence that is derived from an Fc polypeptide; a cytokine targeting polypeptide or a sequence derived from a cytokine targeting polypeptide; a WAP domain containing polypeptide or a sequence derived from a WAP containing polypeptide; and an albumin polypeptide or an amino acid sequence that is derived from a serum albumin polypeptide. This invention also relates to methods of using such molecules in a variety of therapeutic and diagnostic indications, as well as methods of producing such molecules. 153.-. (canceled)54. A method of purifying a fusion protein , the method comprising the steps of:(a) culturing a cell comprising a nucleic acid construct that encodes the fusion protein under conditions that allow for the expression of the fusion protein,wherein the fusion protein comprises at least one human serpin polypeptide comprising an alpha-1 antitrypsin (AAT) polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1, 2, 32, 33, 33, 34, and 35 operably linked to an immunoglobulin Fc polypeptide;(b) contacting a supernatant from the cultured cell with an affinity resin under conditions that allow for binding between the affinity resin and the fusion protein; and(c) eluting the fusion protein from the affinity resin using a buffer under conditions that allow for the detachment of the fusion protein from the affinity resin, wherein the buffer is at a near-neutral pH,wherein the purified fusion protein inhibits neutrophil elastase (NE) activity.55. The method of claim 54 , wherein the cell comprises a Chinese Hamster Ovary (CHO) cell claim 54 , a Human Embryonic Kidney (HEK) 293 cell claim 54 , a COS cell claim 54 , a PER.C6® cell claim 54 , a NS0 ...

Macrocyclic Cysteine Protease Inhibitors and Compositions Thereof

The present invention provides a novel class of macrocyclic compounds, which are useful as cysteine protease inhibitors. Also provided are novel intermediates and methods of preparing the compounds. The invention also provides pharmaceutical compositions comprising the compounds. The compounds and compositions are useful in methods of treating or preventing one or more diseases associated with cysteine protease activity, particularly those associated with calpain activity.

Compositions and Methods of Use for Alpha-1 Antitrypsin Fusion Polypeptides

Embodiments herein report compositions of alpha-1 antitrypsin fusion polypeptides or peptide derivatives thereof. In certain embodiments, compositions and methods relate to generating a construct of use in pharmaceutically acceptable compositions to treat a subject in need of alpha-1 antitrypsin therapy or treatment. In other embodiments, compositions and methods disclosed herein concern linking alpha-1 antitrypsin or derivative thereof to an immune fragment. 1. A method for treating a subject in need of alpha-1 antitrypsin (AAT) therapy comprising , administering to the subject an isolated fusion polypeptide comprising a first polypeptide comprising an AAT polypeptide or a carboxyterminal fragment thereof , wherein the AAT polypeptide or carboxyterminal fragment thereof comprises SEQ ID NO: 1 , SEQ ID NO: 33 , SEQ ID NO: 24 and SEQ ID NO: 25 , SEQ ID NO: 29 , SEQ ID NO: 30 , SEQ ID NO: 31 , SEQ ID NO: 34 , SEQ ID NO: 43 , SEQ ID NO: 44 , SEQ ID NO: 52 , SEQ ID NO: 53 or SEQ ID NO: 54 and a second polypeptide comprising an immunoglobulin Fc polypeptide , wherein the isolated fusion polypeptide is part of a pharmaceutical composition.2. The method according to claim 1 , wherein the first polypeptide of AAT consists of SEQ ID NO:1 or SEQ ID NO:33.3. The method according to claim 1 , wherein the composition is administered by inhalation claim 1 , intranasally claim 1 , intraperitoneally claim 1 , intravaginally claim 1 , orally claim 1 , topically claim 1 , by implant claim 1 , intravenously claim 1 , intramolecularly claim 1 , subcutaneously or by another method of administration.4. The method according to claim 1 , wherein the alpha antitrypsin (AAT) therapy comprises treating AAT deficiency claim 1 , transplant rejection; graft versus host disease (GvHD) claim 1 , a cardiac condition and wherein administering the isolated fusion polypeptide reduces cardiac remodeling; ischemia-reperfusion injury (IR) claim 1 , side effects of reconstructive or plastic surgery; ...

THERAPEUTIC STRATEGIES TO TREAT CNS PATHOLOGY IN MUCOPOLYSACCHARIDOSES

The invention provides for nucleotide sequences encoding for a chimeric sulfatase, viral vectors expressing such sequences for gene therapy and pharmaceutical uses of the chimeric expressed protein. The invention is particularly applied in the therapy of mucopolysaccharidosis, preferably type IIIA. 1. A nucleotide sequence encoding for a chimeric sulfatase , wherein the encoded chimeric sulfatase consists essentially of , in the N-terminal to C-terminal sequence order:a) a signal peptide derived from either the human α-antitrypsin (hAAT) amino acid sequence or the human Iduronate-2-sulfatase (IDS) amino acid sequence;b) a human sulfatase derived amino acid sequence that is deprived of its signal peptide; andc) an ApoB LDLR-binding domain.2. The nucleotide sequence according to claim 1 , wherein the encoded signal peptide has the sequence of SEQ ID NO: 2 or SEQ ID NO: 4.3. The nucleotide sequence according to claim 1 , wherein the encoded human sulfatase derived amino acid sequence of b) claim 1 , is derived from human sulfamidase.4. The nucleotide sequence according to claim 3 , wherein the encoded human sulfamidase derived amino acid sequence consists essentially of the sequence of SEQ ID NO: 8.5. The nucleotide sequence according to claim 1 , wherein the encoded ApoB LDLR-binding domain consists essentially of the sequence of SEQ ID NO: 10.6. The nucleotide sequence according to claim 1 , wherein the nucleotide sequence is selected from the group consisting of:a) Assembly hAATsp-SGSH-3xflag-ApoB cassette (SEQ ID NO: 15),b) Assembly hIDSsp-SGSH-3xflag-ApoB cassette (SEQ ID NO: 17),c) Assembly hAATsp-SGSH-ApoB cassette (SEQ ID NO: 23), andd) Assembly hIDSsp-SGSH-ApoB cassette (SEQ ID NO: 25).7. A recombinant plasmid claim 1 , suitable for gene therapy claim 1 , comprising the nucleotide sequence according to claim 1 , under the control of a liver specific promoter.8. The recombinant plasmid according to claim 7 , wherein the liver specific promoter is the human ...

SERPIN FUSION POLYPEPTIDES AND METHODS OF USE THEREOF

This invention relates to molecules, particularly polypeptides, more particularly fusion proteins that include a serpin polypeptide or an amino acid sequence that is derived from a serpin and second polypeptide comprising of at least one the following: an Fc polypeptide or an amino acid sequence that is derived from an Fc polypeptide; a cytokine targeting polypeptide or a sequence derived from a cytokine targeting polypeptide; a WAP domain containing polypeptide or a sequence derived from a WAP containing polypeptide; and an albumin polypeptide or an amino acid sequence that is derived from a serum albumin polypeptide. This invention also relates to methods of using such molecules in a variety of therapeutic and diagnostic indications, as well as methods of producing such molecules. 114.-. (canceled)15. A method of treating or alleviating a symptom of a disease or disorder associated with aberrant serine protease expression or activity in a subject in need thereof , the method comprising administering a fusion protein comprising at least one human serpin polypeptide operably linked to a modified human immunoglobulin Fc polypeptide , wherein the modified human immunoglobulin Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 60 , wherein the immunoglobulin Fc polypeptide comprises mutations at positions 5228 , L235 , M252 , and M428 , and wherein the human serpin polypeptide is a human alpha-1 antitrypsin (AAT) polypeptide comprising the amino acid sequence of SEQ ID NO: 80.16. A method of treating or alleviating inflammation or a symptom of an inflammatory disease or disorder while reducing the risk of infection , in a subject in need thereof , the method comprising administering a fusion protein comprising at least one human serpin polypeptide operably linked to a modified human immunoglobulin Fc polypeptide , wherein the modified human immunoglobulin Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 60 , wherein the immunoglobulin Fc ...

METHODS FOR ISOLATING BLOOD PRODUCTS FROM AN INTER-ALPHA INHIBITOR PROTEIN-DEPLETED BLOOD PRODUCT MATERIAL

Described is a method for isolating multiple blood products from a single starting material. Isolation of multiple blood products from a single starting material maximizes the efficiency of blood product isolation. In the present invention, one or more blood products are isolated from a blood product material previously depleted of inter-alpha inhibitor protein (IαIp). This method provides new paths for increasing the efficiency of isolating blood components and providing pharmaceutically acceptable forms of those components. 1. A method for isolating one or more blood products from an inter-alpha inhibitor protein (IαIp)-depleted blood product material , comprising:(a) providing an IαIp-depleted blood product material, wherein said IαIp-depleted blood product material is a blood product material depleted of one or more IαIp family members by at least about 20% of the total present in the source blood product material and that includes at least about 20% of IgG present in the source blood product material.(b) isolating one or more blood products from said IαIp-depleted blood product material, wherein at least one of said one or more blood products is selected from albumin, IgA, IgG, IgM, IgD, IgG, IVIg, anti-D IgG, hepatitis B IgG, measles IgG, rabies IgG, tetanus IgG, Varicella Zoster IgG, fibrinogen (factor I), prothrombin (factor II), thrombin, anti-thrombin III, factor III, factor V, factor VII, factor VIII, factor IX, factor X, factor XI, factor XII, factor XIII, fibronectin, alpha-1 antitrypsin, alpha-2 antiplasmin, urokinase, C1-inhibitor, protein C, protein S, protein Z, protein Z-related protease inhibitor, plasminogen, tissue plasminogen activator, plasminogen activator inhibitor-1, plasminogen activator inhibitor-2, von Willebrand factor, factor H, prekallikrein, high-molecular-weight kininogen, and heparin cofactor II.2. The method of claim 1 , wherein said IαIp-depleted blood product material substantially comprises 3 or more non-IαIp blood products ...

MESSENGER UNA MOLECULES AND USES THEREOF

This invention provides a range of translatable messenger UNA (mUNA) molecules. The mUNA molecules can be translated in vitro and in vivo to provide an active polypeptide or protein, or to provide an immunization agent or vaccine component. The mUNA molecules can be used as an active agent to express an active polypeptide or protein in cells or subjects. Among other things, the mUNA molecules are useful in methods for treating rare diseases. 1. A mUNA molecule , comprising one or more UNA monomers , and comprising nucleic acid monomers , wherein the mUNA molecule is translatable to express a polypeptide or protein.2. The molecule of claim 1 , wherein the molecule comprises from 200 to 12 claim 1 ,000 monomers.3. The molecule of claim 1 , wherein the molecule comprises from 200 to 4 claim 1 ,000 monomers.4. The molecule of claim 1 , wherein the molecule comprises from 1 to 8 claim 1 ,000 UNA monomers.5. The molecule of claim 1 , wherein the molecule comprises from 1 to 100 UNA monomers.6. The molecule of claim 1 , wherein the molecule comprises from 1 to 20 UNA monomers.7. The molecule of claim 1 , wherein the molecule comprises one or more modified nucleic acid nucleotides claim 1 , or one or more chemically-modified nucleic acid nucleotides.8. The molecule of claim 1 , wherein the molecule comprises a 5′ cap claim 1 , a 5′ untranslated region of monomers claim 1 , a coding region of monomers claim 1 , a 3′ untranslated region of monomers claim 1 , and a tail region of monomers.9. The molecule of claim 8 , wherein the molecule comprises a translation enhancer in a 5′ or 3′ untranslated region.10. The molecule of claim 1 , wherein the molecule is translatable in vivo.11. The molecule of claim 1 , wherein the molecule is translatable in vitro.12. The molecule of claim 1 , wherein the molecule is translatable in a mammalian cell.13. The molecule of claim 1 , wherein the molecule is translatable in a human in vivo.14. The molecule of claim 1 , wherein a translation ...

BINDING FUSION PROTEINS, BINDING FUSION PROTEIN-DRUG CONJUGATES, XTEN-DRUG CONJUGATES AND METHODS OF MAKING AND USING SAME

The present invention relates to binding fusion protein compositions comprising targeting moieties linked to extended recombinant polypeptide (XTEN), binding fusion protein-drug conjugate compositions, and XTEN-drug conjugate compositions, isolated nucleic acids encoding the compositions and vectors and host cells containing the same, and methods of using such compositions in treatment of diseases, disorders, and conditions. 1. An isolated binding fusion protein comprising a first extended recombinant polypeptide (XTEN) comprising at least 36 amino acid residues and a first targeting moiety with specific binding affinity to a first target , wherein said fusion protein exhibits a terminal half-life that is longer than about 72 hours when administered to a subject.228-. (canceled) This application is a continuation application of U.S. application Ser. No. 15/955,046, filed Apr. 17, 2018 which is a continuation application of U.S. application Ser. No. 14/977,035, filed Dec. 21, 2015, which is a continuation application of U.S. application Ser. No. 13/631,361, filed Sep. 28, 2012 which is a continuvation application of International patent Application PCT/US2011/030992, filed Apr. 1, 2011 which claims the benefit of U.S. Provisional Application Ser. No. 61/341,720 filed Apr. 2, 2010, and 61/341,996 filed Apr. 8, 2010, each of which is incorporated herein by reference in its entirety.This invention was made with government support under SBIR grant 2R44GM079873-02 awarded by the National Institutes of Health. The government has certain rights in the invention.The instant application contains a Sequence Listing which has been submitted in ASCII format via EFS-Web and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Dec. 27, 2012, is named 3280873 Ltd and is 2,247,117 bytes in size.Antibodies or immunoglobulins are molecules that recognize and bind to specific cognate antigens or ligands. Because of their exclusive specificities, antibodies, ...

WAP DOMAIN FUSION POLYPEPTIDES AND METHODS OF USE THEREOF

This invention relates to fusion proteins that include a whey acidic protein (WAP) domain-containing polypeptide and a second polypeptide. Additionally, the invention relates to fusion proteins that include a WAP domain-containing polypeptide a second polypeptide, and a third polypeptide. The second and/or third polypeptides of the fusion proteins of the invention are a Fe polypeptide; an albumin polypeptide; a cytokine targeting polypeptide; or a serpin polypeptide. This invention also relates to methods of using such molecules in a variety of therapeutic and diagnostic indications, as well as methods of producing such molecules. 1. An isolated fusion protein comprising at least one human whey acidic protein (WAP) domain-containing polypeptide operably linked to a second polypeptide , wherein the second polypeptide comprises at least one the following:an immunoglobulin Fc polypeptide or an amino acid sequence that is derived from an immunoglobulin Fc polypeptide;a cytokine targeting polypeptide or a sequence derived from a cytokine targeting polypeptide;a serpin polypeptide or a sequence derived from a serpin polypeptide; oran albumin polypeptide or an amino acid sequence that is derived from a serum albumin polypeptide.2. An isolated fusion protein comprising at least one human secretory leukocyte proteinase inhibitor (SLPI) polypeptide operably linked to a second polypeptide , wherein the second polypeptide comprises at least one of the following:an immunoglobulin Fc polypeptide;a cytokine targeting polypeptide;an alpha-1 antitrypsin (AAT) polypeptide;an alpha-1 antitrypsin (AAT) polypeptide and an immunoglobulin Fc polypeptide; anda human albumin (HSA) polypeptide.3. An isolated fusion protein comprising human at least one human Elafin polypeptide operably linked to a second polypeptide , wherein the second polypeptide comprises at least one of the following:an immunoglobulin Fc polypeptide;a cytokine targeting polypeptide;an alpha-1 antitrypsin (AAT) ...

SERPIN FUSION POLYPEPTIDES AND METHODS OF USE THEREOF

This invention relates to molecules, particularly polypeptides, more particularly fusion proteins that include a serpin polypeptide or an amino acid sequence that is derived from a serpin and second polypeptide comprising of at least one the following: an Fc polypeptide or an amino acid sequence that is derived from an Fc polypeptide; a cytokine targeting polypeptide or a sequence derived from a cytokine targeting polypeptide; a WAP domain containing polypeptide or a sequence derived from a WAP containing polypeptide; and an albumin polypeptide or an amino acid sequence that is derived from a serum albumin polypeptide. This invention also relates to methods of using such molecules in a variety of therapeutic and diagnostic indications, as well as methods of producing such molecules. 1. An isolated fusion protein comprising at least one human serpin polypeptide operably linked to a second polypeptide wherein the second polypeptide comprises at least one the following:an immunoglobulin Fc polypeptide or an amino acid sequence that is derived from an immunoglobulin Fc polypeptide;a cytokine targeting polypeptide or a sequence derived from a cytokine targeting polypeptide;a WAP domain containing polypeptide or a sequence derived from a WAP domain containing polypeptide; oran albumin polypeptide or an amino acid sequence that is derived from a serum albumin polypeptide.2. An isolated fusion protein comprising at least one alpha-1 antitrypsin (AAT) polypeptide operably linked to a second polypeptide of at least one of the following:an immunoglobulin Fc polypeptide;a cytokine targeting peptide;a human secretory leukocyte proteinase inhibitor (SLPI) polypeptide; andan immunoglobulin Fc polypeptide and a human SLPI polypeptide, wherein the SLPI polypeptide comprises the WAP2 domain of human SLPI; ora human serum albumin (HSA) polypeptide.3. An isolated fusion protein comprising at least one alpha-1 antitrypsin (AAT) polypeptide operably linked to a second polypeptide of at ...

MUTANT ALPHA-1-ANTITRYPSIN COMPOSITIONS AND USE THEREOF

The present invention provides mutant alpha 1-antitrypsin proteins, pharmaceutical compositions comprising the same, and methods of use thereof in treatment of subjects with an inflammatory disease or disorder. 1. An isolated polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 1 or a fragment , a derivative or an analog thereof , wherein X is any amino acid other than cysteine.2. The isolated polypeptide of claim 1 , wherein said analog comprises an amino acid sequence with at least 70% homology to SEQ ID NO: 2.3. The isolated polypeptide of claim 1 , wherein X is selected from the group consisting of: proline claim 1 , valine claim 1 , threonine claim 1 , serine and isoleucine.4. The isolated polypeptide of claim 3 , wherein X is proline.5. The isolated polypeptide of claim 1 , further comprising a mutation at position 357 claim 1 , wherein a proline at position 357 is mutated to any amino acid other than proline.6. The isolated polypeptide of claim 5 , wherein said proline is mutated to an amino acid selected from the group consisting of: cysteine claim 5 , alanine claim 5 , methionine claim 5 , isoleucine claim 5 , and valine.7. The isolated polypeptide of claim 6 , said proline is mutated to an amino acid selected from cysteine and alanine.8. The isolated polypeptide of claim 1 , wherein said polypeptide has at least one therapeutic property that is greater than said therapeutic property of recombinant human alpha 1-antitrypsin (rhAAT) protein (SEQ ID NO: 2) or serum purified human alpha 1-antitrypsin (hAAT).9. The isolated polypeptide of claim 8 , wherein said therapeutic property is selected from an anti-inflammatory property and a wound healing property.10. The isolated polypeptide of claim 9 , wherein said anti-inflammatory property is selected from reducing secretion of a pro-inflammatory cytokine and reducing activation of macrophages.11. The isolated polypeptide of claim 10 , wherein said pro-inflammatory cytokine is selected from IL-6 ...

SERUM-FREE STABLE TRANSFECTION AND PRODUCTION OF RECOMBINANT HUMAN PROTEINS IN HUMAN CELL LINES

The present invention relates to an improved method for the serum-free production of an immortalized human cell line stably transfected under serum-free conditions with a specific vector carrying the gene coding for the protein of interest. Furthermore the invention relates to a production cell line obtained by said method, a production method for said protein of interest utilizing said production cell line, and the specific vector carrying the gene of interest itself. 117-. (canceled)18. A transfection vector comprising (a) an origin of replication , (b) a nucleic acid sequence encoding a human plasma protein selected from a blood clotting factor , a growth factor , a colony-stimulating factor (CSFs) , a cytokine , an immunoglobulin , a protease , a protease inhibitor , a transport protein , a hormone , an inhibitory or regulatory acting protein , and derivatives and mutants thereof , (c) a promoter , and (d) a bovine growth hormone polyadenylation (poly (A)) signal.19. The vector of claim 18 , wherein(i) the promoter is selected from a viral promoter, a housekeeping gene promoter, and a tissue specific promoter; and/or(ii) the origin of replication allows the replication and amplification of the plasmid in bacteria; and/or(iii) the vector further carries at least one gene for a selection marker and/or is under control of the promoter as defined in (i) above; and/or(iv) the vector further carries one or more further regulatory elements.20. The vector of claim 18 , wherein said promoter and poly(A) signal are linked to the 5′ and 3′ end of the gene encoding said human target protein claim 18 , respectively.21. The vector of claim 18 , wherein said human plasma protein is selected from factor IX as encoded by bps 939 to 2324 of SEQ ID NO: 1 claim 18 , human A1AT as encoded by bps 913 to 2259 of SEQ ID NO: 2 claim 18 , wild-type factor VIII as shown in SEQ ID NO: 9 claim 18 , B-domain deleted human factor VIII as encoded by bps 783 to 5162 of SEQ ID NO: 3 claim 18 , ...

CHEMICALLY INDUCIBLE CUCUMBER MOSAIC VIRUS PROTEIN EXPRESSION SYSTEM

The invention relates to a novel chemically inducible plant viral amplicon (CMViva) expression system that permits controllable, high level expression of foreign genes in plant hosts. This system employs agro-infiltration of plants to provide a transient production of a protein of interest, such as a human blood protein. This system provides a major advantage over existing plant expression systems because it allows for consistent expression of foreign or heterologous proteins in plant hosts. 1. A plant expression system comprising a cucumber mosaic virus inducible viral amplicon for the expression of a heterologous gene in a plant or plant cell culture.2. The expression system of claim 1 , wherein heterologous gene codes for a human protein.3. A plant or plant cell comprising the expression system of .4. The expression system of claim 1 , wherein the expression system further comprises a cDNA encoding a CMV1a replicase and a cDNA encoding a CMV2a replicase.5. The expression system of claim 4 , wherein said cDNA encoding said CMV1a replicase is operably linked to a chemically-inducible LEX operator and an estradiol inducible expression system.6. The expression system of claim 4 , wherein said cDNA encoding said CMV2a replicase is operably linked to a CaM 35S promoter.7. The expression system of claim 1 , wherein the heterologous gene is operably linked to a CaM 35S promoter.8. The expression system of further comprising a cDNA encoding a gene silencing suppressor.9. A method for producing a heterologous protein claim 1 , said method comprising:{'claim-ref': [{'@idref': 'CLM-00001', 'claims 1'}, {'@idref': 'CLM-00008', '8'}], 'a) contacting a plant, excised plant tissue, or plant cell with a bacterial cell containing the expression system of -;'}b) contacting the plant, excised plant tissue, or culturing the plant cell with a chemical inducer;c) growing the contacted plant, culturing the plant cell, orincubating the excised plant tissue for a time period sufficient to ...

PROTEIN AND METABOLITE ENRICHMENT USING FOCUSED ACOUSTIC ENERGY

Apparatus and method for disassociating protein complexes, e.g., to allow recovery and/or analysis of at least one of the proteins or metabolites released from a complex. Disassociation is done using focused acoustic energy and without solvents, excessive heat or other process conditions that damage proteins or metabolites. Disassociation may be followed by depletion of one of the proteins released from complexes, e.g., to allow another protein or metabolite released from the complexes to be recovered. 1. A method of analyzing proteins and/or metabolites in a sample , comprising:providing a sample including a plurality of different types of protein, at least two of the types of protein forming a plurality of complexes in which a first protein is bound to a second protein; andexposing the sample to focused acoustic energy to disrupt the plurality of complexes and disassociate the first protein from the second protein in each of the complexes.2. The method of claim 1 , wherein the second protein has a higher molecular weight than the first protein.3. The method of claim 1 , wherein the first protein is sequestered at least partially within the second protein prior to disassociation from the second protein.4. The method of claim 1 , wherein the disassociation of the first and second proteins is achieved at a sample temperature below 60 degrees C.5. The method of claim 1 , further comprising depleting the sample of the second protein.6. The method of claim 5 , further comprising recovering and identifying the first protein from the sample after depletion of the second protein from the sample.7. The method of claim 1 , wherein the sample includes blood plasma claim 1 , and the first protein is present in the sample at a first concentration that is at least an order of magnitude lower than a second concentration at which the second protein is present in the sample.8. The method of claim 7 , wherein the first protein is present in the sample free of any complex with the ...

PEPTIDES AND METHODS OF USING THE SAME

We describe peptides and their uses for the treatment of autoimmune, inflammatory and metabolic diseases. 115.-. (canceled)16. A composition comprising a peptide selected from the group consisting of: X1 is V or L;', 'X2 is V, L or M;', 'X3 is M, I or V;', 'Z1 is any amino acid;', 'Z2 is a sequence of any two amino acids; and', 'Z3 is a sequence any five amino acids, and wherein the peptide is 37 or fewer amino acids;, '(a) a peptide consisting of the amino acid sequence X1-Z1-F-N-K-P-F-X2-Z2-X3-Z3-Q (SEQ ID NO: 2), wherein'}(b) a peptide consisting of the amino acid sequence VKFNKPFVFLMIEQNTK (SEQ ID NO: 1); X1 is V or L;', 'X2 is K or R;', 'X3 is V, L or M;', 'X4 is M, I or V;', 'X5 is K or Q;', 'Z1 is any amino acid;', 'Z2 is a sequence of any two amino acids; and', 'Z3 is a sequence any five amino acids;, '(c) a peptide consisting essentially of the amino acid sequence X1-Z1-F-N-X2-P-F-X3-Z2-X4-Z3-X5 (SEQ ID NO: 3), wherein'}(d) a peptide consisting essentially of the amino acid sequence RFNRPFLR (SEQ ID NO: 4).(e) a peptide consisting essentially of the amino acid sequence of RRRFNRPFLRRR (SEQ ID NO: 8).(f) a peptide consisting essentially of the amino acid sequence of VKFNKPFVFLMIEQNTK (SEQ ID NO: 1); and(g) a peptide consisting essentially of the amino acid sequence of FNRPFL (SEQ ID NO: 10).171. The composition of claim , wherein the peptide further comprises at least one second peptide or protein.18. The composition of claim 17 , wherein the at least one second protein or peptide is attached to the peptide as a fusion peptide.19. The composition of claim 17 , wherein the at least one second peptide or protein is an epitope tag or a half-life extender or both.201. The composition of claim claim 17 , wherein the peptide comprises one or more D-amino acids.211. The composition of claim claim 17 , wherein the peptide consists of 37 amino acid residues or fewer.221. The composition of claim claim 17 , wherein the peptide consists of 35 amino acid residues or ...

METHODS AND COMPOSITIONS FOR MODULATING ALPHA-1-ANTITRYPSIN EXPRESSION

Disclosed herein are methods for decreasing A1AT mRNA and protein expression and treating, ameliorating, preventing, slowing progression, or stopping progression of fibrosis. Disclosed herein are methods for decreasing A1AT mRNA and protein expression and treating, ameliorating, preventing, slowing progression, or stopping progression of liver disease, such as, A1ATD associated liver disease, and pulmonary disease, such as, A1ATD associated pulmonary disease in an individual in need thereof. Methods for inhibiting A1AT mRNA and protein expression can also be used as a prophylactic treatment to prevent individuals at risk for developing a liver disease, such as, A1ATD associated liver disease and pulmonary disease, such as, A1ATD associated pulmonary disease. 13-. (canceled)4. A compound comprising a modified oligonucleotide consisting of 20 to 30 linked nucleosides and having a nucleobase sequence comprising a portion of at least 20 contiguous nucleobases that is 100% complementary to an equal length portion of nucleobases 1349 to 1597 of SEQ ID NO: 1 , and wherein the nucleobase sequence of the modified oligonucleotide is at least 90% complementary to SEQ ID NO: 1 , and wherein each nucleotide of the modified oligonucleotide is a modified nucleotide having , independently , a modified sugar moiety , a modified internucleoside linkage , and/or modified nucleobase.523-. (canceled)24. A composition comprising a compound according to and a pharmaceutically acceptable carrier or diluent.25. (canceled)26. A method of treating claim 24 , ameliorating or preventing an A1AT deficiency disease in an animal comprising administering to an animal having or at risk of developing an A1AT deficiency disease a composition according to claim 24 , and thereby treating claim 24 , ameliorating claim 24 , or preventing the A1AT deficiency disease in the animal.2749-. (canceled) The present application is being filed along with a Sequence Listing in electronic format. The Sequence ...

Serum-free Stable Transfection and Production of Recombinant Human Proteins in Human Cell Lines

The present invention relates to an improved method for the serum-free production of an immortalized human cell line stably transfected under serum-free conditions with a specific vector carrying the gene coding for the protein of interest. Furthermore the invention relates to a production cell line obtained by said method, a production method for said protein of interest utilizing said production cell line, and the specific vector carrying the gene of interest itself.

Mouse Model of Alpha-One Antitrypsin (AAT) Deficiency

Transgenic non-human animals, e.g., rodents, e.g., mice comprising genomic mutations that inactive all of the serpin1A genes and thus lack any functional serpinA1 genes. As a result of the genomic mutations, the animals express no hepatic or circulatory AAT protein. Also provided herein are cells and tissues derived from the transgenic mice. 1. A method of evaluating an effect of a test compound on development of Alpha-One Antitrypsin (AAT)-deficiency lung disease , the method comprising:providing a transgenic mouse whose genome comprises inactivating mutations in exon 2 of each of Serpina1a, Serpina1b, Serpina1c, Serpina1d, and Serpina1e genes, and which expresses no hepatic or circulatory AAT protein;exposing the mouse to the test compound; andevaluating a parameter of respiratory physiology in the transgenic mouse.2. The method of claim 1 , wherein the inactivating mutations comprise deletions in exon 2.3. The method of claim 1 , wherein the transgenic mouse is a mouse model of Alpha-One Antitrypsin (AAT)-deficiency lung disease.4. The method of claim 1 , wherein the test compound is an environmental factor.5. The method of claim 4 , wherein the environmental factor is a toxin.6. The method of claim 1 , wherein the mouse is exposed to untested air particulates or vaping.7. The method of claim 1 , comprising measuring the parameter in the animal before exposure claim 1 , and during and/or after exposure to the test compound.8. The method of claim 1 , wherein the parameter of respiratory physiology is respiratory volume claim 1 , inspiratory capacity claim 1 , elastance claim 1 , compliance claim 1 , and/or quasi-static compliance.9. The method of claim 1 , further comprising:exposing the mouse to a potential therapeutic; andevaluating a parameter of respiratory physiology in the transgenic mouse.10. The method of claim 9 , comprising exposing the mouse to a potential therapeutic before claim 9 , during and/or after exposing the mouse to the test compound.11. A ...

COMPOSITIONS, METHODS AND USES FOR ALPHA-1 ANTITRYPSIN OR DERIVATIVES THEREOF AS RADIOPROTECTANTS

Embodiments herein report methods and compositions for treating or preventing adverse effects of radiation therapies. In certain embodiments, compositions and methods relate to reducing or inhibiting damage due to acute, periodic or chronic radiation exposure. 1. A method for ameliorating adverse effects of radiation exposure in a subject comprising , administering to a subject undergoing or scheduled to undergo radiation therapy a therapeutically effective amount of a composition comprising alpha-1 antitrypsin (AAT) , a fragment thereof or a mutant thereof wherein the composition modulates the adverse effects of the radiation therapy in the subject.2. The method of claim 1 , wherein the composition comprises naturally occurring AAT (SEQ ID NO:1 or SEQ ID NO:39).3. The method of claim 1 , wherein the composition comprises a composition of one or more carboxyterminal fragments of naturally occurring AAT.4. The method of claim 1 , wherein the subject is undergoing radiation therapy for a condition.5. The method of claim 4 , wherein the condition is cancer.6. The method of claim 1 , wherein the subject has been exposed to radiation as a result of a nuclear accident claim 1 , nuclear test or nuclear attack.7. The method of claim 1 , wherein the subject has undergone claim 1 , or is undergoing a diagnostic procedure that comprises exposure to radiation.8. The method of claim 1 , wherein the composition is administered in a therapeutically effective amount to reduce or prevent at least one radiation-induced effect selected from the group consisting of myelosuppression claim 1 , renal toxicity claim 1 , weight loss claim 1 , behavioral changes claim 1 , pain claim 1 , nausea claim 1 , vomiting claim 1 , diarrhea claim 1 , constipation claim 1 , all malnutrition claim 1 , hair loss claim 1 , numbness claim 1 , changes in tastes claim 1 , loss of appetite claim 1 , thinned or brittle hair claim 1 , mouth sores claim 1 , memory loss claim 1 , hemorrhage claim 1 , ...

Raav-based compositions and methods for treating alpha-1 anti-trypsin deficiencies

The invention relates to isolated nucleic acids and rAAV-based compositions, methods and kits useful for treating genetic diseases (e.g., alpha-1 antitrypsin deficiency).

Lipid Nanoparticle Compositions and Methods for mRNA Delivery

Disclosed herein are compositions and methods for modulating the production of a protein in a target cell. The compositions and methods disclosed herein are capable of ameliorating diseases associated with protein or enzyme deficiencies. 140-. (canceled)41. A method for delivery of messenger RNA (mRNA) for in vivo production of a protein , comprising administering , to a human , a composition comprising an mRNA that encodes the protein wherein the mRNA is encapsulated within a lipid nanoparticle , wherein the administering of the composition results in expression of the protein encoded by the mRNA that is detectable in serum at least 72 hours after administration , and wherein the lipid nanoparticle comprises one or more PEG-modified lipids.42. The method of claim 41 , wherein the mRNA comprises a 5′ untranslated region.43. The method of claim 42 , wherein the mRNA comprises a 3′ untranslated region.44. The method of claim 43 , wherein the mRNA comprises a cap structure.45. The method of claim 44 , wherein the mRNA comprises a poly A tail.46. The method of claim 45 , wherein the lipid nanoparticle comprises one or more cationic lipids.47. The method of claim 46 , wherein the lipid nanoparticle comprises one or more non-cationic lipids.48. The method of claim 47 , wherein the mRNA is unmodified.49. The method of claim 48 , wherein the composition is a reconstituted lyophilized composition.50. The method of claim 47 , wherein the modification comprises a modified nucleotide.51. The method of claim 50 , wherein the modified nucleotide is pseudouridine.52. The method of claim 51 , wherein the composition is a reconstituted lyophilized composition.53. The method of claim 48 , wherein the composition is administered by intravenous injection.54. The method of claim 51 , wherein the composition is administered by intravenous injection.55. The method of claim 48 , wherein the composition is administered by intramuscular injection.56. The method of claim 51 , wherein the ...

SERPIN FUSION POLYPEPTIDES AND METHODS OF USE THEREOF

This invention relates to molecules, particularly polypeptides, more particularly fusion proteins that include a serpin polypeptide or an amino acid sequence that is derived from a serpin and second polypeptide comprising of at least one the following: an Fc polypeptide or an amino acid sequence that is derived from an Fc polypeptide; a cytokine targeting polypeptide or a sequence derived from a cytokine targeting polypeptide; a WAP domain containing polypeptide or a sequence derived from a WAP containing polypeptide; and an albumin polypeptide or an amino acid sequence that is derived from a serum albumin polypeptide. This invention also relates to methods of using such molecules in a variety of therapeutic and diagnostic indications, as well as methods of producing such molecules. 153.-. (canceled)54. An isolated fusion protein for inhibiting serine proteases having at least one human alpha-1 antitrypsin (AAT) polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO: 1 , SEQ ID NO: 2 , SEQ ID NO: 32 , and SEQ ID NO: 33 , wherein the AAT polypeptide is operably linked to an immunoglobulin Fc polypeptide having an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO: 6.55. The isolated fusion protein of claim 54 , wherein the AAT polypeptide and the immunoglobulin Fc polypeptide are operably linked via a hinge region claim 54 , a linker region claim 54 , or both a hinge region and linker region.56. The isolated fusion protein of claim 55 , wherein the peptide sequence comprises the amino acid sequence of SEQ ID NO: 43 claim 55 , SEQ ID NO: 44 claim 55 , SEQ ID NO: 45 claim 55 , or SEQ ID NO: 46.57. The isolated fusion protein of claim 54 , wherein the human AAT polypeptide comprises of the amino acid sequence of SEQ ID NO: 1 operably linked to an immunoglobulin Fc polypeptide comprising the amino acid sequence of SEQ ID NO: 6.58. The isolated fusion protein of claim 54 , wherein the human AAT ...

METHOD, COMPOSITION, AND ARTICLE OF MANUFACTURE FOR PROVIDING ALPHA-1 ANTITRYPSIN

The present invention provides a method for providing alpha-1 antitrypsin (α1-AT) to a subject, in particular a method for treating or preventing a disorder or disease associated with α1-AT deficiency in the subject, wherein the method comprises providing, subcutaneously, a therapeutically or prophylactically effective amount of α1-AT to the subject. Also provided is a composition and article of manufacture comprising α1-AT, in particular a formulation suitable for subcutaneous administration of α1-AT. 1. A method for treating a disorder or disease associated with α1-AT deficiency in a subject , the method comprising:administering subcutaneously to the subject in need thereof a dose of α1-AT, wherein the dose of α1-AT administered is about 120% of a dose of α1-AT administered to the subject by intravenous route; wherein the dose administered by intravenous route achieves a blood α1-AT trough level of at least about 80 mg/dL;wherein the amount administered to the subject by intravenous route is therapeutically effective in preventing or treating a disorder or disease associated with α1-AT deficiency; andwherein the frequency of α1-AT subcutaneous administration is sufficient to maintain a trough level of at least about 80 mg/dL.2. The method of claim 1 , wherein the α1-AT is a plasma-derived α1-AT.3. The method of claim 1 , wherein the dose of α1-AT is administered in combination withone or more reagents comprising a hyaluronidase.4. The method of claim 3 , wherein the subcutaneous administration of α1-AT achieves at least about a 1.40-fold higher plasma Crelative to an identical subcutaneous administration of α1-AT without one or more reagents comprising hyaluronidase.5. The method of claim 4 , wherein the subcutaneous administration of α1-AT achieves at least about a 2-fold lower Trelative to an identical subcutaneous administration without one or more reagents comprising hyaluronidase.6. The method of claim 1 , wherein the subcutaneous administration of α1-AT does ...

Methods and compositions for single chain variable region enox2 antibodies for cancer detection and diagnosis

Cancers of different cellular or tissue origins express different ENOX2 cancer isoforms or combinations of isoforms and shed these proteins into the circulation. Herein are disclosed methods both for cancer detection and diagnosis of particular origin, based on the patterns and molecular weights of the isoforms which allow the identification of the cell type and or tissue of origin of the neoplasm. Relative ENOX2 amounts are proportional to tumor burden and provide a reliable measure of response to therapy and disease progression. Also provided is the amino acid sequence to which the scFv antibodies bind as the molecular basis for the specificity of the test.

ALPHA1 -ANTITRYPSIN COMPOSITIONS AND METHODS OF TREATING AUTOIMMUNE DISEASES

The specification provides compositions comprising chimeric proteins comprising AAT conjugated to an Fc region of an immunoglobulin. Methods for treating autoimmune disease, e.g., diabetes, e.g., Type 1 and Type 2 diabetes, are also provided. 1. A recombinant polypeptide comprising an α1-antitrypsin polypeptide (AAT) conjugated to an Fc region of an immunoglobulin.2. The recombinant polypeptide of claim 1 , wherein the AAT comprises an amino acid sequence that is at least 90% identical to SEQ ID NO:2.3. The recombinant polypeptide of claim 1 , wherein the immunoglobulin is selected from the group consisting of IgG claim 1 , IgA claim 1 , IgD claim 1 , IgE claim 1 , and IgM.4. The recombinant polypeptide of claim 1 , wherein the immunoglobulin is selected from the group consisting of IgG1 claim 1 , IgG2 claim 1 , IgG3 claim 1 , and IgG4.5. The recombinant polypeptide of claim 1 , wherein the Fc region of the immunoglobulin comprises an amino acid sequence that is at least 90% identical to SEQ ID NO:4 or 6.6. The recombinant polypeptide of claim 1 , wherein polypeptide comprises an amino acid sequence that is at least 90% identical to SEQ ID NO:8 claim 1 , 10 claim 1 , 12 claim 1 , 14 claim 1 , 16 claim 1 , or 18.7. (canceled)8. A nucleic acid molecule encoding a recombinant polypeptide comprising an AAT conjugated to an Fc region of an immunoglobulin.9. The nucleic acid molecule of claim 8 , wherein the AAT comprises an amino acid sequence that is least 90% identical to SEQ ID NO:2.10. The nucleic acid molecule of claim 8 , wherein the nucleic acid sequence encoding AAT is at least 90% identical to SEQ ID NO:1.11. The nucleic acid molecule of claim 8 , wherein the nucleic acid sequence encoding the Fc region of the immunoglobulin is at least 90% identical to SEQ ID NO:3 or 5.12. The nucleic acid molecule of claim 8 , wherein the nucleic acid molecule comprises a nucleic acid sequence is at least 90% identical to SEQ ID NO:7 claim 8 , 9 claim 8 , 11 claim 8 , 13 claim ...

ADENO-ASSOCIATED VIRUS SEROTYPE I NUCLEIC ACID SEQUENCES, VECTORS AND HOST CELLS CONTAINING SAME

The nucleic acid sequences of adeno-associated virus (AAV) serotype 1 are provided, as are vectors and host cells containing these sequences and functional fragments thereof. Also provided are methods of delivering genes via AAV-1 derived vectors. 1. (canceled)2. A composition comprising 1×10to 1×10genomes of a recombinant adenovirus having an AAV1 capsid comprising a vp1 protein , a vp2 protein , and a vp3 protein , wherein said vp3 protein has an amino acid sequence of SEQ ID NO:17 , wherein said recombinant virus further comprises a heterologous molecule which comprises an AAV 5′ inverted terminal repeat sequence (ITR) , a transgene , and an AAV 3′ ITR , and a pharmaceutically acceptable carrier.3. The composition according to claim 2 , comprising 1×10to 1×10genomes.4. The composition according to claim 1 , wherein said vp1 protein has the amino acid sequence of SEQ ID NO: 13.5. The composition according to claim 1 , wherein said vp2 protein has the amino acid sequence of SEQ ID NO: 15.6. The composition according to claim 1 , wherein the 5′ ITR and 3′ ITR are of AAV serotype 2.7. The composition according to claim 1 , further comprising a promoter which directs expression of the transgene.8. The composition according to claim 1 , wherein said transgene encodes a protein or peptide.9. The composition according to claim 8 , wherein said protein or peptide is a therapeutic protein or peptide.10. The composition according to claim 8 , wherein said protein or peptide is an immunogenic protein or peptide.11. The composition according to claim 1 , wherein said transgene encodes a cytokine claim 1 , a hormone claim 1 , or a growth factor.12. The composition according to claim 1 , wherein said composition is formulated for delivery to muscle.13. The composition according to claim 7 , wherein the promoter is a cytomegalovirus promoter. This is a continuation of U.S. patent application Ser. No. 14/849,722, filed Sep. 10, 2015, which is continuation of U.S. patent ...

LIPID NANOPARTICLE COMPOSITIONS AND METHODS FOR MRNA DELIVERY

Disclosed herein are compositions and methods for modulating the production of a protein in a target cell. The compositions and methods disclosed herein are capable of ameliorating diseases associated with protein or enzyme deficiencies. 140-. (canceled)41. A method for delivery of messenger RNA (mRNA) for in vivo production of a secreted polypeptide , comprising administering , to a human , a composition comprising an mRNA that encodes the secreted polypeptide wherein the mRNA is encapsulated within a lipid nanoparticle , wherein the administering of the composition results in expression of the secreted polypeptide encoded by the mRNA that is detectable in serum at least 72 hours after administration , and wherein the lipid nanoparticle comprises one or more PEG-modified lipids.42. The method of claim 41 , wherein the mRNA comprises a 5′ untranslated region.43. The method of claim 42 , wherein the mRNA comprises a 3′ untranslated region.44. The method of claim 43 , wherein the mRNA comprises a cap structure.45. The method of claim 44 , wherein the mRNA comprises a poly A tail.46. The method of claim 45 , wherein the lipid nanoparticle comprises one or more cationic lipids.47. The method of claim 46 , wherein the lipid nanoparticle comprises one or more non-cationic lipids.48. The method of claim 47 , wherein the mRNA is unmodified.49. The method of claim 48 , wherein the composition is a reconstituted lyophilized composition.50. The method of claim 47 , wherein the modification comprises a modified nucleotide.51. The method of claim 50 , wherein the modified nucleotide is pseudouridine.52. The method of claim 51 , wherein the composition is a reconstituted lyophilized composition.53. The method of claim 48 , wherein the composition is administered by intravenous injection.54. The method of claim 51 , wherein the composition is administered by intravenous injection.55. The method of claim 48 , wherein the composition is administered by intramuscular injection.56. ...

Methods of treatment using alpha-1-antitrypsin compositions

A streamlined method for purifying alpha-1-antitrypsin (AAT) from an AAT-containing protein mixture, such as Coh fraction IV precipitate, is provided. In the method of the invention, contaminating proteins are destabilized by cleavage of disulfide bonds with a reducing reagent, such as dithiol, which does not affect AAT. The destabilized proteins are then preferentially adsorbed on a solid protein-adsorbing material, without the addition of a salt as a precipitante. Spearation of the solid absorbent from the solution leaves a purified AAT solution that is directly suitabale for chromatographic purification, without the need for extensive desalting as in prior art processes. A process incorporating this method, which provides pharmaceutical grade AAT in high yield on a commercial scale, is also described.

Compositions, methods and uses for alpha-1 antitrypsin fusion molecules

Embodiments herein report compositions of alpha-1 antitrypsin fusion polypeptides or peptide derivatives thereof. In certain embodiments, compositions and methods relate to generating a construct of use in pharmaceutically acceptable compositions to treat a subject in need of alpha-1 antitrypsin therapy or treatment. In other embodiments, compositions and methods disclosed herein concern linking alpha-1 antitrypsin or derivative thereof to an immune fragment.

Ethanol dependence of alpha1 antitrypsin c-terminal lys truncation by basic carboxypeptidases

The present invention provides methods of preparing alpha-1-antiproteinase inhibitor and controlling the amount of des-lys alpha-1-antiproteinase inhibitor in the preparation, and compositions comprising the same, as well as methods of treatment using the same.

RAAV-BASED COMPOSITIONS AND METHODS

The invention relates to isolated nucleic acids and rAAV-based compositions, methods and kits useful for treating genetic diseases (e.g., alpha-1 antitrypsin deficiency). 1. An isolated nucleic acid comprising:(a) a first region that encodes one or more first miRNAs comprising a nucleic acid having sufficient sequence complementary with an endogenous mRNA of a subject to hybridize with and inhibit expression of the endogenous mRNA, wherein the endogenous mRNA encodes a first protein; and(b) a second region encoding an exogenous mRNA that encodes a second protein, wherein the second protein has an amino acid sequence that is at least 85% identical to the first protein,wherein the one or more first miRNAs do not comprise a nucleic acid having sufficient sequence complementary to hybridize with and inhibit expression of the exogenous mRNA, and wherein the first region is positioned within an untranslated portion of the second region.2. The isolated nucleic acid of claim 1 , wherein the untranslated portion is an intron.3. The isolated nucleic acid of claim 1 , wherein the first region is between the first codon of the exogenous mRNA and 1000 nucleotides upstream of the first codon.4. An isolated nucleic acid comprising:(a) a first region encoding one or more first miRNAs comprising a nucleic acid having sufficient sequence complementary with an endogenous mRNA of a subject to hybridize with and inhibit expression of the endogenous mRNA, wherein the endogenous mRNA encodes a first protein; and(b) a second region encoding an exogenous mRNA that encodes a second protein, wherein the second protein has an amino acid sequence that is at least 85% identical to the first protein,wherein the one or more first miRNAs do not comprise a nucleic acid having sufficient sequence complementary to hybridize with and inhibit expression of the exogenous mRNA, and wherein the first region is positioned downstream of a portion of the second region encoding the poly-A tail of the exogenous ...

Pharmaceutical Composition Containing Fusion Protein and Use Thereof

This disclosure is directed to a fusion protein composition comprising an alpha-1-antitrypsin or α1-antitrypsin (also known as A1AT, A1A, or AAT) polypeptide (AAT), a modified AAT (mAAT) or a functional variant thereof and a bioactive polypeptide. This disclosure is particularly directed to a pharmaceutical composition comprising the fusion protein for treating a disease, such as a cancer or an autoimmune disease. The bioactive polypeptide can be a peptide hormone, interferon, or cytokine, such as interleukin-2 (IL-2), a modified IL-2 (mIL-2), IL-15, G-CSF, GM-CSF, IFN-α2, IFN-β1, GLP-1, FGF21, sdAb, a fragment thereof, a modified polypeptide thereof, or a combination thereof. One advantage of the fusion protein is to enhance the activity, stability, bioavailability or a combination thereof, of the bioactive polypeptide. 1. A fusion protein composition comprising an AAT polypeptide or a functional variant thereof , and a bioactive polypeptide , wherein said bioactive polypeptide is covalently linked to said AAT polypeptide , covalently linked to said AAT polypeptide via a linker peptide , or a combination thereof;wherein said AAT polypeptide comprises a mAAT polypeptide or a functional variant thereof, wherein said mAAT polypeptide or said functional variant thereof is free from cysteine amino acid residue, wherein said functional variant has at least 85% sequence identity of said mAAT polypeptide and wherein said mAAT polypeptide and said functional variant each is free from serine protease inhibitor activity.2. The fusion protein composition of claim 1 , wherein said fusion protein composition comprises said linker peptide that has an N-terminal claim 1 , a C-terminal and 1-50 amino acid residues and wherein said linker peptide is positioned between said AAT polypeptide and said bioactive polypeptide.3. The fusion protein composition of claim 2 , wherein said bioactive polypeptide is linked to the N-terminal of said linker peptide and said AAT polypeptide is ...

PHARMACEUTICAL COMPOSITIONS CONTAINING POLYPEPTIDES DERIVED FROM ALPHA-1 ANTITRYPSIN AND METHODS OF USE THEREOF

The present invention relates to isolated polypeptides comprising the amino acid sequence of residues 378-413 of α-1-antitrypsyn (serpina1c), and active fragments thereof, and to pharmaceutical compositions comprising same. The compositions of the invention are useful for treating burns, inflammatory, autoimmune and degenerative diseases. 1. A composition comprising an isolated polypeptide , wherein said composition is pharmaceutically acceptable , said polypeptide diminishes or abrogates an inflammatory response in a subject and said polypeptide shares at least 95% identity with a polypeptide consisting of an amino acid sequence selected from the group consisting of SEQ ID NO:1 , SEQ ID NO:2 , SEQ ID NO: 3 and SEQ ID NO: 4.2. The composition according to claim 1 , wherein said amino acid sequence shares at least 97% identity with that set forth in SEQ ID NO: 1.3. The composition according to claim 1 , wherein said amino acid sequence shares at least 99% identity with that set forth in SEQ ID NO: 1.4. The composition according to claim 1 , wherein said amino acid sequence consists of that set forth in SEQ ID NO: 1.5. The composition according to claim 1 , wherein said sequence shares at least 97% identity with that set forth in SEQ ID NO:2.6. The composition according to claim 1 , wherein said sequence shares at least 99% identity with that set forth in SEQ ID NO:2.7. The composition according to claim 1 , wherein said sequence consists of that set forth in SEQ ID NO:2.8. The composition according to claim 1 , wherein said sequence shares at least 97% identity with that set forth in SEQ ID NO: 3 or 4.9. The composition according to claim 1 , wherein said sequence shares at least 99% identity with that set forth in SEQ ID NO: 3 or 4.10. The composition according to claim 1 , wherein said sequence consists of that set forth in SEQ ID NO: 3 or 4.1110. The composition according to any one of - claim 1 , wherein said polypeptide is a fragment of the polypeptide of .12. A ...

ALPHA-1-ANTITRYPSIN COMPOSITIONS

A streamlined method for purifying alpha-1-antitrypsin (AAT) from an AAT-containing protein mixture, such as a Cohn fraction IV precipitate, is provided. In the method of the invention, contaminating proteins are destabilized by cleavage of disulfide bonds with a reducing reagent, such as a dithiol, which does not affect AAT. The destabilized proteins are then preferentially adsorbed on a solid protein-adsorbing material, without the addition of a salt as a precipitant. Separation of the solid adsorbent from the solution leaves a purified AAT solution that is directly suitable for chromatographic purification, without the need for extensive desalting as in prior art processes. A process incorporating this method, which provides pharmaceutical grade AAT in high yield on a commercial scale, is also described. 117.-. (canceled)18. A pharmaceutical composition , comprising alpha-1-antitrypsin (AAT) isolated or purified from plasma or fractions thereof , wherein said composition further comprises:(a) less than 0.1% albumin; and(b) less than 0.1% apolipoprotein A1, and{'sub': '1 cm,280 nm', 'sup': '1%', 'wherein percentages of albumin and apolipoprotein A1 are weight percentages based on total protein weight, and wherein total protein weight is measured using a Bradford assay or by absorbance at 280 nm using as an extinction coefficient E=5.3.'}19. The composition of claim 18 , further comprising:(c) less than 0.1% transferrin, andwherein percentages of albumin, apolipoprotein A1, and transferrin are weight percentages based on total protein weight.20. The composition of claim 18 , wherein the AAT is pasteurized and/or subjected to a viral reduction procedure.21. The composition of claim 20 , wherein the AAT is subjected to a viral reduction procedure capable of reducing the number of enveloped viruses by at least 11 logunits and capable of reducing the number of non-enveloped viruses by at least 6 logunits.22. The composition of claim 20 , wherein the viral reduction ...

METHODS AND COMPOSITIONS FOR MODULATING ALPHA-1-ANTITRYPSIN EXPRESSION

Disclosed herein are methods for decreasing A1AT mRNA and protein expression and treating, ameliorating, preventing, slowing progression, or stopping progression of fibrosis. Disclosed herein are methods for decreasing A1AT mRNA and protein expression and treating, ameliorating, preventing, slowing progression, or stopping progression of liver disease, such as, A1ATD associated liver disease, and pulmonary disease, such as, A1ATD associated pulmonary disease in an individual in need thereof. Methods for inhibiting A1AT mRNA and protein expression can also be used as a prophylactic treatment to prevent individuals at risk for developing a liver disease, such as, A1ATD associated liver disease and pulmonary disease, such as, A1ATD associated pulmonary disease. 13-. (canceled)4. A compound comprising a modified oligonucleotide consisting of 20 to 30 linked nucleosides and having a nucleobase sequence comprising a portion of at least 20 contiguous nucleobases that is 100% complementary to an equal length portion of nucleobases 1349 to 1597 of SEQ ID NO: 1 , and wherein the nucleobase sequence of the modified oligonucleotide is at least 90% complementary to SEQ ID NO: 1 , and wherein the modified oligonucleotide comprises at least one modified nucleoside comprising a modified sugar.58-. (canceled)9. The compound of claim 4 , wherein the modified oligonucleotide consists of 20 or 22 linked nucleosides.10. The compound of claim 4 , wherein the nucleobase sequence of the modified oligonucleotide is 100% complementary to SEQ ID NO: 1.11. The compound of claim 4 , wherein the modified oligonucleotide is single-stranded.12. The compound of claim 4 , wherein at least one internucleoside linkage of the modified oligonucleotide is a modified internucleoside linkage.13. The compound of claim 12 , wherein at least one modified internucleoside linkage is a phosphorothioate linkage.14. The compound of claim 13 , wherein each internucleoside linkage is a phosphorothioate ...

COMPOSITIONS, METHODS AND USES FOR ALPHA-1 ANTITRYPSIN FUSION MOLECULES

Embodiments herein report compositions of and methods for making and using alpha-1 antitrypsin (AAT) fusion molecules or peptide derivatives thereof. In certain embodiments, compositions and methods relate to generating an AAT fusion molecule of use in pharmaceutically acceptable compositions to treat a subject in need of AAT therapy or treatment. In other embodiments, compositions and methods disclosed herein concern linking AAT or derivative thereof to an immune fragment. 1. A construct comprising:a nucleic acid encoding an alpha-1 antitrypsin (AAT) polypeptide or fragment or peptide cleavage molecule thereof; anda nucleic acid encoding an immunoglobulin Fc fragment, the nucleic acid further manipulated to truncate or eliminate the immune fragment hinge region.2. The construct of claim 1 , wherein the nucleic acid encoding AAT comprises a nucleic acid encoding naturally occurring AAT (SEQ ID NO:1).3. The construct of wherein the nucleic acid encoding the AAT peptide cleavage molecule comprises a nucleic acid encoding one or more carboxyterminal fragments cleavage products of naturally occurring AAT.4. The construct of claim 3 , wherein the carboxyterminal fragment comprises the last 80 amino acids of SEQ ID NO:1 or 10 or more consecutive amino acids thereof5. The construct of claim 1 , wherein the immunoglobulin Fc comprises a nucleic acid encoding an Fc fragment from IgG1 claim 1 , IgG2 claim 1 , IgG3 or IgG4.6. The construct of claim 1 , wherein the nucleic acid encoding AAT or fragment thereof comprises a nucleic acid represented by SEQ ID NO: 47 claim 1 , SEQ ID NO: 48 claim 1 , SEQ ID NO: 50 claim 1 , SEQ ID NO: 52 claim 1 , or SEQ ID NO: 54.7. The construct of claim 1 , wherein the encoded AAT polypeptide is represented by the amino acid sequence of SEQ ID NO: 54.8. The construct of claim 1 , wherein the nucleic acid encoding the AAT or fragment or peptide cleavage molecule thereof comprises a nucleic acid encoding a mutant having a mutation at the reactive ...

COMPOSITIONS OF, AND METHODS FOR, ALPHA-1 ANTITRYPSIN Fc FUSION MOLECULES

A novel method of treating and preventing bacterial diseases is provided. In particular, the present invention relates to compositions and methods for inhibition of Gram negative, Gram positive and acid fast bacilli in general and (TB), complex (MAC), and anthrax in particular. Thus, the invention relates to modulation of cellular activities, including macrophage activity, and the like. More particularly, the present invention relates to the inhibitory compounds comprising naturally occurring and man-made inhibitors of serine protease. 1. A construct comprising a nucleic acid sequence encoding a fusion polypeptidecomprising:a first nucleic acid sequence encoding mammalian alpha-1 antitrypsin (AAT), wherein the first nucleic acid sequence comprises SEQ ID NO: 68 or SEQ ID NO:78; anda second nucleic acid sequence encoding an IgG1 Fc.2. The construct according to claim 1 , wherein the construct comprises an encoded fusion protein of mammalian alpha-1 antitrypsin (AAT) fused to the amino-terminus or carboxy-terminus of Fc.3. The construct according to claim 1 , wherein the encoded fusion polypeptide is part of a pharmaceutical composition that further comprises a pharmaceutically acceptable carrier or excipient.4. The construct according to claim 1 , wherein the second nucleic acid sequence encoding the IgG1 Fc is represented by SEQ ID NO: 64.5. The construct according to claim 1 , wherein the construct encoding a fusion polypeptide comprises the nucleic acid sequence represented by SEQ ID NO: 86 or SEQ ID NO: 88.6. The construct according to claim 1 , wherein the construct encodes a fusion polypeptide represented by SEQ ID NO: 87 or SEQ ID NO: 89.7. A vector comprising the nucleic acid construct of .8. An expression vector comprising the nucleic acid construct of and a promoter operatively linked thereto.9. A transformed cell comprising the expression vector of .10. An isolated preparation of expressed inclusion bodies comprising an encoded fusion polypeptide of the ...

Diagnosis Of Sepsis And Systemic Inflammatory Response Syndrome

The present invention relates a method for the diagnosis, prediction or risk stratification for mortality and/or disease outcome of a subject that has or is suspected to have sepsis, comprising determining the presence and/or level of antitrypsin (ATT) or fragments thereof in a sample taken from said subject and/or determining the presence and/or level of transthyretin (TTR) or fragments thereof, wherein the presence and/or level of ATT and/or TTR or fragments thereof is correlated with an increased risk of mortality and, wherein said increased risk of mortality and/or poor disease outcome is given if the level of ATT is below a certain cut-off value and/or the level of fragments thereof is above a certain cut-off value and/or said increased risk of mortality and/or poor disease outcome is given if the level of TTR is below a certain cut-off value and/or the level of fragments thereof is below a certain cut-off value. The invention relates in general to the use of ATT and/or TTR or its fragments for the diagnosis of sepsis, and to nucleotides of SEQ ID NO. 2 to 14. 1. A method for the diagnosis , prediction or risk stratification for mortality or disease outcome of a subject that has or is suspected to have sepsis , comprising the steps of (a)-(c):(a) determining the level of antitrypsin (ATT) or fragments thereof in a sample taken from said subject,(b) wherein the level of ATT or fragments thereof is correlated with an increased risk of mortality or poor disease outcome and, wherein 'and/or comprising the steps of (d)-(f):', '(c) said increased risk of mortality or poor disease outcome is given if the level of ATT is below a certain cut-off value and/or the level of fragments thereof is above a certain cut-off value;'}(d) determining the level of transthyretin (TTR) or fragments thereof in a sample taken from said subject,(e) wherein the level of TTR or fragments thereof is correlated with an increased risk of mortality or poor disease outcome and, wherein(f) said ...

Compositions, methods and uses for alpha-1 antitrypsin fusion molecules

Embodiments herein report compositions of alpha-1 antitrypsin fusion polypeptides or peptide derivatives thereof. In certain embodiments, compositions and methods relate to generating a construct of use in pharmaceutically acceptable compositions to treat a subject in need of alpha-1 antitrypsin therapy or treatment. In other embodiments, compositions and methods disclosed herein concern linking alpha-1 antitrypsin or derivative thereof to an immune fragment.

COMPOSITIONS, METHODS AND USES FOR ALPHA-1 ANTITRYPSIN FUSION MOLECULES

Embodiments herein report compositions of and methods for making and using alpha-1 antitrypsin (AAT) fusion molecules or peptide derivatives thereof. In certain embodiments, compositions and methods relate to generating an AAT fusion molecule of use in pharmaceutically acceptable compositions to treat a subject in need of AAT therapy or treatment. In other embodiments, compositions and methods disclosed herein concern linking AAT or derivative thereof to an immune fragment. 1. A nucleic acid construct encoding a fusion polypeptide represented by SEQ ID NO: 49 , SEQ ID NO: 56 , SEQ ID NO: 57 , or SEQ ID NO: 58.2. A nucleic acid construct encoding the fusion polypeptide represented by SEQ ID NO: 49.3. A composition comprising a fusion polypeptide of and a pharmaceutically acceptable carrier.4. A vector comprising the nucleic acid construct of .5. An isolated preparation of expressed inclusion bodies comprising a fusion polypeptide of .6. A nucleic acid construct encoding a fusion polypeptide represented by SEQ ID NO: 56.7. A composition comprising a fusion polypeptide of and a pharmaceutically acceptable carrier.8. A vector comprising the nucleic acid construct of .9. An isolated preparation of expressed inclusion bodies comprising a fusion polypeptide of .10. A composition comprising a fusion polypeptide of and a pharmaceutically acceptable carrier.11. A vector comprising the nucleic acid construct of .12. An isolated preparation of expressed inclusion bodies comprising a fusion polypeptide of .13. A nucleic acid construct encoding the fusion polypeptide represented by SEQ ID NO: 57.14. A composition comprising a fusion polypeptide of and a pharmaceutically acceptable carrier.15. A vector comprising the nucleic acid construct of .16. An isolated preparation of expressed inclusion bodies comprising a fusion polypeptide of .17. A nucleic acid construct encoding a fusion polypeptide represented by SEQ ID NO: 58.18. A composition comprising a fusion polypeptide of and a ...

RAAV-BASED COMPOSITIONS AND METHODS

The invention relates to isolated nucleic acids and rAAV-based compositions, methods and kits useful for treating genetic diseases (e.g., alpha-1 antitrypsin deficiency). 1. A recombinant adenoviral associated vector (rAAV vector) comprising:(a) a first region that encodes one or more first miRNAs comprising a nucleic acid having sufficient sequence complementary with an endogenous mRNA of a subject to hybridize with and inhibit expression of the endogenous mRNA, wherein the endogenous mRNA encodes a first protein having a dominant negative or gain of function mutation; and(b) a second region encoding an exogenous mRNA that encodes a second protein, wherein the second protein has an amino acid sequence that is at least 85% identical to the first protein and does not have the dominant negative or gain of function mutation,wherein the one or more first miRNAs do not comprise a nucleic acid having sufficient sequence complementary to hybridize with and inhibit expression of the exogenous mRNA, and wherein the first region is positioned within between a portion of the second region encoding the last codon of the exogenous mRNA and a polyadenylation sequence of the exogenous mRNA.24-. (canceled)5. The rAAV vector of claim 1 , further comprising a third region encoding a one or more second miRNAs comprising a nucleic acid having sufficient sequence complementary to hybridize with and inhibit expression of the endogenous mRNA claim 1 , wherein the third region is positioned within an untranslated portion of the second region.67-. (canceled)8. The rAAV vector of claim 5 , wherein the third region is between the first codon of the exogenous mRNA and a position 1000 nucleotides upstream of the first codon.9. The rAAV vector of claim 1 , wherein the first region encodes two first miRNAs.10. The rAAV vector of claim 1 , wherein the first region encodes three first miRNAs.11. The rAAV vector of claim 5 , wherein the third region encodes two second miRNAs.12. The rAAV vector of ...

MODIFIED SERPINS FOR THE TREATMENT OF BLEEDING DISORDERS

This invention relates pro-coagulant serpin molecules engineered by modification of the P4, P2, P1 and/or P1′ residues within the reactive center loop (RCL) to display increased specificity for anticoagulant proteases. These modified serpin molecules may be useful in therapy, for example as pro-coagulants for the treatment of bleeding. 1. A modified serpin having mutations at residues P1′ and P2 and optionally P1 and/or P4 in the reactive center loop (RCL) thereof ,wherein the P1′ residue is mutated to Q, H, K or R; and the P2 residue is mutated to H, K or R, and;wherein said mutations increase the inhibition of one or more anticoagulant proteases selected from activated Protein C and thrombin:thrombomodulin complex relative to the inhibition of one or more procoagulant proteases selected from thrombin, fVIIa, fXa, fIXa and fXIa.2. A modified serpin according to wherein the P1′ residue is mutated to K or Q.3. A modified serpin according to wherein the P2 residue is mutated to K.4. A modified serpin according to wherein the P2 and the P1′ residues respectively in the modified serpin are KK claim 1 , RK claim 1 , RH claim 1 , KH claim 1 , RQ or KQ.5. A modified serpin according to wherein the modified serpin comprises a mutation at P4 claim 1 , optionally wherein the P4 residue is mutated to F claim 1 , S claim 1 , R claim 1 , V claim 1 , C claim 1 , W claim 1 , K claim 1 , G claim 1 , L claim 1 , H claim 1 , T claim 1 , Q or A.6. A modified serpin according to wherein the modified serpin comprises a mutation at P1 claim 1 , optionally wherein the P1 residue is mutated to R.7. A modified serpin according to wherein the modified serpin has an R residue at the P1 position.8. A modified serpin according to wherein the mutations in the RCL of the modified serpin consist of mutations at positions P1′ and P2; positions P1′ claim 1 , P2 and P1; positions P1′ claim 1 , P2 and P4; or positions P1′ claim 1 , P2 claim 1 , P4 and P1.9. A modified serpin according to wherein;the ...

COMPOSITIONS OF, AND METHODS FOR, ALPHA-1 ANTITRYPSIN FcFUSION MOLECULES

A novel method of treating and preventing bacterial diseases is provided. In particular, the present invention relates to compositions and methods for inhibition of Gram negative, Gram positive and acid fast bacilli in general and tuberculosis (TB), complex (MAC), and anthrax in particular. Thus, the invention relates to modulation of cellular activities, including macrophage activity, and the like. More particularly, the present invention relates to the inhibitory compounds comprising naturally occurring and man-made inhibitors of serine protease. 1. A pharmaceutical composition of fusion polypeptide construct comprising:a single alpha-1 antitrypsin fusion polypeptide comprising:a first polypeptide comprising mammalian alpha-1 antitrypsin (AAT); anda second polypeptide comprising an immunoglobulin constant region.2. The construct of claim 1 , wherein the first polypeptide comprising AAT further comprises a signal sequence.3. A transformed cell comprising a nucleic acid construct encoding the construct of .4. An isolated preparation of expressed inclusion bodies comprising the construct of .5. A process for producing a construct comprising culturing the transformed cell of claim 3 , so that the construct is expressed claim 3 , and recovering the expressed construct.6. A method for treating a subject in need of AAT therapy claim 2 , the method comprising administering to the subject a therapeutically effective amount of the composition of in an amount effective to treat the subject.7. A method for treating a subject having an infection from a pathogenic organism claim 1 , the method comprising administering to the subject a therapeutically effective amount of the composition of in an amount effective to treat the infection.8. The pharmaceutical composition of wherein the composition is formulated for inhalation claim 2 , intranasal claim 2 , intravenous claim 2 , intramolecular or subcutaneous administration.9. The pharmaceutical composition of claim 1 , wherein the ...

NOVEL ALPHA-1 ANTITRYPSIN VARIANT, PREPARATION METHOD THEREOF AND USE THEREOF

A novel alpha-1 antitrypsin variant, a method of preparing the same, and use thereof are provided. The alpha-1 antitrypsin variant has excellent stability in the body and maintains an inhibitory effect on elastase activities because the blood half-life (t) and the area under blood drug concentration vs. time curve (AUC) are remarkably increased by adding an N-glycosylation site in animal cells through amino acid mutation between 1and 25positions of the N-terminus of alpha-1 antitrypsin. Therefore, the alpha-1 antitrypsin variant can be useful in preventing or treating alpha-1 antitrypsin deficiency. 1. An alpha-1 antitrypsin variant prepared by substituting an amino acid at a specific site between 1and 25positions of the N-terminus of alpha-1 antitrypsin in order to add a glycosylation site.2. The alpha-1 antitrypsin variant of claim 1 , wherein the alpha-1 antitrypsin variant has 1 to 3 glycosylation sites added thereto.3. The alpha-1 antitrypsin variant of claim 1 , wherein the specific site is present between 3and 13positions of the N-terminus.4. The alpha-1 antitrypsin variant of claim 1 , wherein the specific site is present at a 9or 12position of the N-terminus.5. The alpha-1 antitrypsin variant of claim 1 , wherein the specific sites are present at 4and 9positions claim 1 , 4and 12positions claim 1 , or 9and 12positions.6. A method of preparing an alpha-1 antitrypsin variant of claim 1 , comprising:{'sup': st', 'th, 'a) substituting an amino acid at a specific site between 1and 25positions of the N-terminus of alpha-1 antitrypsin in order to add a glycosylation site;'}b) culturing cells transformed with an alpha-1 antitrypsin expression vector having the glycosylation site added thereto in a culture medium;c) expressing an alpha-1 antitrypsin variant protein from the cells; andd) purifying and recovering the expressed alpha-1 antitrypsin variant protein.7. A composition for preventing or treating alpha-1 antitrypsin deficiency claim 1 , comprising an alpha-1 ...

COMPOSITIONS AND METHODS FOR ALPHA-1-ANTITRYPSIN DISORDERS

Disclosed herein are compositions and methods useful for treating an alpha-1-antitrypsin deficiency. 1. A recombinant protein comprising:(a) an alpha-1-antitrypsin serpin domain; and(b) a human serum albumin binding domain.2. The recombinant protein according to claim 1 , further comprising a linker consisting of SEQ ID NO: 63.3. The recombinant protein of any one of the preceding claims claim 1 , wherein the C-terminus of the alpha-1-antitrypsin serpin domain is fused to the N-terminus of the human serum albumin binding domain.4. The recombinant protein of any one of the preceding claims claim 1 , wherein the human serum albumin binding domain is at least a portion of a human serum albumin having either SEQ ID NO: 3 or SEQ ID NO: 5.5. The recombinant protein of any one of the preceding claims claim 1 , wherein half-life of the alpha-1-antitrypsin serpin domain compared to wild type alpha-1-antitrypsin is increased.6. The recombinant protein of any one of the preceding claims claim 1 , wherein a number of N-glycans or polysialyation of the recombinant protein in the alpha-1-antitrypsin serpin domain is greater than a wild type alpha-1-antitrypsin serpin domain.7. The recombinant protein according to any one of the preceding claims claim 1 , wherein the alpha-1-antitrypsin serpin domain has one or more mutations as compared to a wild type alpha-1-antitrypsin serpin domain.8. The recombinant protein of claim 7 , wherein the one or more mutations causes resistance to methionine oxidation as compared to a wild type alpha-1-antitrypsin serpin domain.9. The recombinant protein of or claim 7 , wherein the alpha-1-antitrypsin serpin domain retains activity compared to a wild type alpha-1-antitrypsin serpin domain.10. The recombinant protein of or claim 7 , wherein the one or more mutations comprise M351V and/or M358V with residues being numbered according to SEQ ID NO: 1.11. The recombinant protein of any one of - claim 7 , wherein the one or more mutations comprise K168C ...

ALPHA1 -ANTITRYPSIN COMPOSITIONS AND METHODS OF TREATING AUTOIMMUNE DISEASES

The specification provides compositions comprising chimeric proteins comprising AAT conjugated to an Fc region of an immunoglobulin. Methods for treating autoimmune disease, e.g., diabetes, e.g., Type 1 and Type 2 diabetes, are also provided. 1. A recombinant polypeptide comprising an al-antitrypsin polypeptide (AAT) conjugated to an Fc region of an immunoglobulin.2. The recombinant polypeptide of claim 1 , wherein the AAT comprises an amino acid sequence that is at least 90% identical to SEQ ID NO:2.3. The recombinant polypeptide of claim 1 , wherein the immunoglobulin is selected from the group consisting of IgG IgA claim 1 , IgD claim 1 , IgE claim 1 , and IgM.4. The recombinant polypeptide of claim 1 , wherein the immunoglobulin is selected from the group consisting of IgG1 claim 1 , IgG2 claim 1 , IgG3 claim 1 , and IgG4.5. The recombinant polypeptide of claim 1 , wherein the Fc region of the immunoglobulin comprises an amino acid sequence that is at least 90% identical to SEQ ID NO:4 or 6.6. The recombinant polypeptide of claim 1 , wherein polypeptide comprises an amino acid sequence that is at least 90% identical to SEQ ID NO:8 claim 1 , 10 claim 1 , 12 claim 1 , 14 claim 1 , 16 claim 1 , or 18.7. (canceled)8. A nucleic acid molecule encoding a recombinant polypeptide comprising an AAT conjugated to an Fc region of an immunoglobulin.9. The nucleic acid molecule of claim 8 , wherein the AAT comprises an amino acid sequence that is least 90% identical to SEQ ID NO:2.10. The nucleic acid molecule of claim 8 , wherein the nucleic acid sequence encoding AAT is at least 90% identical to SEQ ID NO:1.11. The nucleic acid molecule of claim 8 , wherein the nucleic acid sequence encoding the Fc region of the immunoglobulin is at least 90% identical to SEQ ID NO:3 or 5.12. The nucleic acid molecule of claim 8 , wherein the nucleic acid molecule comprises a nucleic acid sequence is at least 90% identical to SEQ ID NO:7 claim 8 , 9 claim 8 , 11 claim 8 , 13 claim 8 , 15 ...

PROTEIN EXPRESSION ENHANCING POLYPEPTIDES

Fusion proteins comprising a protein expression enhancing polypeptide linked to a target protein binding domain and nucleic acid molecules encoding such fusion proteins are described for use in enhancing expression and/or location of a targeted protein of interest, for restoring lost functions in cells, and for treating disease. Additional fusion proteins comprising a target protein of interest modified with a fusion partner comprising a protein expression enhancing polypeptide are also disclosed. 2. The polypeptide according to claim 1 , wherein the protein expression enhancing polypeptide is an isolated J domain of a J protein.3. The polypeptide according to claim 2 , wherein the isolated J domain is an isolated J domain of an ERdj protein claim 2 , a large T antigen of SV40 claim 2 , or a mammalian cysteine string protein alpha (CSP-α).4. The polypeptide according to claim 3 , wherein the isolated J domain is an isolated J domain of an Erdj protein selected from Erdj1 claim 3 , Erdj2 claim 3 , Erdj3 claim 3 , Erdj4 claim 3 , Erdj5 claim 3 , Erdj6 and Erdj7.5. The polypeptide according to claim 4 , wherein the isolated J domain is an isolated J domain of Erdj3.6. The polypeptide according to claim 1 , wherein the protein expression enhancing polypeptide is an active fragment of a J domain.8. The polypeptide according to claim 1 , wherein the protein expression enhancing polypeptide is a J domain analog polypeptide claim 1 , wherein said J domain analog polypeptide comprises the amino acid sequence of formula I.9. The polypeptide according to claim 8 , wherein said J domain analog polypeptide comprises the amino acid sequence (SEQ ID NO:326) wherein:X1 is I, L, V, A, or M;the dipeptide X2-X3 is selected from the group consisting of: KR, KK, RK, RR, AK, AR, KA, IK, NK, KQ, RQ, and RD;X4 is A, S, T, R, Q, E, F, C, or I;X5 is Y or F;the dipeptide X6-X7 is selected from the group consisting of: KR, KK, RK, RR, RQ, FR, RL, KL, HK, LK, QK, and KV; andthe dipeptide X8-X9 ...

PURIFICATION OF CELL CULTURE DERIVED ALPHA1 PROTEASE INHIBITOR

Described herein are methods for purifying recombinant, cell culture derived alpha-protease inhibitor and removing a colored species that co-purifies with the recAPI protein. Also described are methods for reducing the iron in cell culture derived alpha-protease inhibitor. 1. A method of purifying cell culture derived human A1PI from an aqueous solution comprising recA1PI , comprising:(a) performing a viral inactivation step on a solution containing recA1PI;(b) passing the virally inactivated solution through an anion exchanger so that recA1PI binds to anion exchanger;(c) eluting recA1PI from the anion exchange resin to obtain an anion exchange eluate containing recA1PI;(d) adding a reducing agent to the anion exchange eluate containing recA1PI to obtain a reducing solution;(e) incubating the reducing solution;(f) passing the reducing solution through a hydrophobic interaction chromatography (HIC) resin so that recA1PI binds to the HIC resin; and(g) eluting recA1PI from the HIC resin to obtain an HIC eluate that contains recA1PI.2. The method of claim 1 , wherein the viral inactivation comprises a solvent/detergent incubation.3. The method of claim 2 , wherein the solvent is added in a range of 0.01% to about 0.5%.4. The method of claim 2 , wherein the detergent is added from about 0.5% to about 2.0% weight per volume of the resulting mixture.5. The method of claim 2 , wherein the solvent/detergent incubation comprises adding about 0.5% polysorbate 20 and about 0.03% tri(n-butyl phosphate) at pH of about 8 and a temperature of about 30° C.6. The method of claim 1 , wherein the anion exchanger is a quaternary ammonium resin.7. The method of claim 6 , wherein the quaternary ammonium resin is Capto™ Q.8. The method of claim 1 , wherein the reducing agent is cysteine (Cys); cysteamine; dithiothreitol (DTT); dithioerythritol (DTE); glutathione (GSH); 2-mercaptoethanol (2-ME); 2-mercaptoethylamine (2-MEA); tris(2-carboxyethyl)phosphine (TCEP); oxalic acid; formic acid; ...

MODIFIED SERPINS FOR THE TREATMENT OF BLEEDING DISORDERS

This invention relates pro-coagulant serpin molecules engineered by modification of the P4, P2, P1 and/or P1′ residues within the reactive center loop (RCL) to display increased specificity for anticoagulant proteases. These modified serpin molecules may be useful in therapy, for example as pro-coagulants for the treatment of bleeding. 1. A modified serpin having mutations at one or both of residues P1′ and P2 and optionally P1 and/or P4 in the reactive center loop (RCL) thereof.2. A modified serpin according to wherein said mutations increase the inhibition of one or more of anticoagulant proteases selected from activated Protein C and thrombin:thrombomodulin complex relative to the inhibition of one or more procoagulant proteases selected from thrombin claim 1 , fVIIa claim 1 , fXa claim 1 , fIXa and fXIa.3. A modified serpin according to or wherein said mutations increase the inhibition of activated Protein C relative to the inhibition of thrombin.4. A modified serpin according to any one of the preceding claims wherein the RCL of the modified serpin consists of the amino acid sequence of the RCL of a wild-type serpin with said mutations therein.5. A modified serpin according to any one of the preceding claims having a mutation at the P1′ position.6. A modified serpin according to wherein the P1′ residue is mutated to Q claim 5 , N claim 5 , Y claim 5 , W claim 5 , R claim 5 , H claim 5 , K claim 5 , C claim 5 , A claim 5 , I claim 5 , T claim 5 , S claim 5 , M claim 5 , P claim 5 , E or V.7. A modified serpin according to wherein the P1′ residue is mutated to Q claim 6 , N claim 6 , Y claim 6 , R claim 6 , H claim 6 , K claim 6 , C claim 6 , A claim 6 , I claim 6 , S claim 6 , M claim 6 , E or V.8. A modified serpin according to wherein the P1′ residue is mutated to Q claim 7 , H claim 7 , K or R.9. A modified serpin according to wherein the P1′ residue is mutated to K.10. A modified serpin according to wherein the P1′ residue is mutated to Q.11. A modified ...

Processes for purifying proteins from plasma

The invention provides processes for producing preparations (e.g., plasma preparations or fibrinogen (Fg)-depleted preparations) containing one or more proteins (e.g., plasma proteins). Processes of the invention can be used to obtain enriched preparations of one or more proteins (e.g., Fg, immunoglobulin (Ig; e.g., IgG), alpha-1 proteinase inhibitor (A1 PI), albumin, plasminogen, prothrombin complex, and/or other plasma proteins). Multiple enriched preparations can be obtained from a single sample (e.g., a whole blood or plasma sample) using the processes of the invention.

METHODS FOR EXTRACTING PROTEINS FROM BLOOD PLASMA

A method of producing protein products including alpha-1-proteinase inhibitor, gamma globulin, albumin, and other proteins from plasma includes steps of: (1) adding a salt to the blood product to produce a first intermediate, wherein the salt comprises between 11-13 wt % of the first intermediate; (2) separating the first intermediate to produce a first supernatant and a first paste; (3) adding a salt to the first intermediate to produce a second intermediate, wherein the salt comprises between 21-23 wt % of the second intermediate; (4) separating the second intermediate to produce a second supernatant and a second paste; (5) separating a third intermediate from the second supernatant by affinity chromatography; and (6) separating the third intermediate by ion exchange chromatography to produce an eluate containing the protein product. Advantageously, the inventive methods are simple and produce alpha-1-proteinase inhibitor, gamma globulin, albumin, and other proteins in high yields. 123.-. (canceled)24. A method of producing a protein product from a blood plasma containing product , the method comprising:adding a salt to the blood plasma containing product to produce a first intermediate, wherein the salt comprises between 11-20 wt % of the first intermediate;separating the first intermediate to produce a first supernatant and a first paste;adding a salt to the first supernatant to produce a second intermediate, wherein the salt comprises between 15-30 wt % of the second intermediate;separating the second intermediate to produce a second supernatant and a second paste;separating a third intermediate from the second supernatant by affinity chromatography; andseparating the third intermediate by ion exchange chromatography to produce an eluate containing the protein product.25. The method of claim 24 , further comprising thawing and stirring the blood plasma containing product prior to adding the salt to the blood product to produce the first intermediate.26. The ...

RAAV-BASED COMPOSITIONS AND METHODS FOR TREATING ALPHA-1 ANTI-TRYPSIN DEFICIENCIES

The invention relates to isolated nucleic acids and rAAV-based compositions, methods and kits useful for treating genetic diseases (e.g., alpha-1 antitrypsin deficiency). 1. A recombinant adenoviral associated vector (rAAV vector) comprising:(a) a first region that encodes one or more first miRNAs comprising a nucleic acid having sufficient sequence complementary with an endogenous mRNA of a subject to hybridize with and inhibit expression of the endogenous mRNA, wherein the endogenous mRNA encodes a first protein having a dominant negative or gain of function mutation; and(b) a second region encoding an exogenous mRNA that encodes a second protein, wherein the second protein has an amino acid sequence that is at least 85% identical to the first protein and does not have the dominant negative or gain of function mutation,wherein the one or more first miRNAs do not comprise a nucleic acid having sufficient sequence complementary to hybridize with and inhibit expression of the exogenous mRNA, and wherein the first region is positioned within between a portion of the second region encoding the last codon of the exogenous mRNA and a polyadenylation sequence of the exogenous mRNA.24-. (canceled)5. The rAAV vector of claim 11 , further comprising a third region encoding a one or more second miRNAs comprising a nucleic acid having sufficient sequence complementary to hybridize with and inhibit expression of the endogenous mRNA claim 11 , wherein the third region is positioned within an untranslated portion of the second region.67-. (canceled)8. The rAAV vector of claim 5 , wherein the third region is between the first codon of the exogenous mRNA and a position 1000 nucleotides upstream of the first codon.9. The rAAV vector of claim 1 , wherein the first region encodes two first miRNAs.10. The rAAV vector of claim 1 , wherein the first region encodes three first miRNAs.11. The rAAV vector of claim 5 , wherein the third region encodes two second miRNAs.12. The rAAV vector of ...

Mouse Model of Alpha-One Antitrypsin (AAT) Deficiency

Transgenic non-human animals, e.g., rodents, e.g., mice comprising genomic mutations that inactive all of the serpinlA genes and thus lack any functional serpinA1 genes. As a result of the genomic mutations, the animals express no hepatic or circulatory AAT protein. Also provided herein are cells and tissues derived from the transgenic mice. 1. A transgenic animal whose genome comprises inactivating mutations in Serpina1a , Serpina1b , Serpina1c , Serpina1d , and Serpina1e genes , and which expresses no hepatic or circulatory Alpha-One Antitrypsin (AAT) protein.2. The transgenic animal of claim 1 , which is a mouse.3. The transgenic animal of claim 1 , wherein the inactivating mutations are in exon 2 of each of the genes.4. The transgenic animal of claim 3 , wherein the inactivating mutations comprise deletions in exon 2.5. The transgenic animal of claim 1 , which is a mouse model of Alpha-One Antitrypsin (AAT)-deficiency lung disease.6. An isolated cell claim 1 , tissue claim 1 , or organ from the transgenic animal of .7. A method of identifying a candidate compound for the treatment of Alpha-One Antitrypsin (AAT)-deficiency lung disease claim 1 , the method comprising:{'claim-ref': [{'@idref': 'CLM-00001', 'claim 1'}, {'@idref': 'CLM-00001', 'claim 1'}], 'contacting the transgenic animal of or a cell, tissue, or organ from the transgenic animal of , with a test compound;'}measuring levels of elastase in the animal, cell, tissue, or organ in the presence and absence of the test compound; andidentifying a test compound that increases levels of elastase as a candidate compound.8. A method of identifying a candidate therapeutic compound for the treatment of Alpha-One Antitrypsin (AAT)-deficiency lung disease claim 1 , the method comprising:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, 'exposing the transgenic animal of to a test compound;'}measuring one or more parameters of respiratory physiology in the animal in the presence and absence of the test compound; ...

VECTORS WITH PROMOTER AND ENHANCER COMBINATIONS FOR TREATING PHENYLKETONURIA

A lentiviral vector system for expressing a lentiviral particle is disclosed. The lentiviral vector system includes a therapeutic vector. The lentiviral vector system produces a lentiviral particle for upregulating PAH expression in the cells of a subject afflicted with phenylketonuria (PKU). 1. A viral vector comprising a therapeutic cargo portion , wherein the therapeutic cargo portion comprises:a PAH sequence or a variant thereof,a promoter; anda liver-specific enhancer,wherein the PAH sequence or the variant thereof is operatively controlled by both the promoter and the liver-specific enhancer.2. The viral vector of claim 1 , wherein the liver-specific enhancer comprises a prothrombin enhancer.3. The viral vector of claim 2 , wherein the promoter comprises a liver-specific promoter.4. The viral vector of claim 3 , wherein the liver-specific promoter comprises a hAAT promoter.5. The viral vector of claim 1 , wherein the PAH sequence or the variant thereof is truncated.6. The viral vector of claim 5 , wherein the PAH sequence or the variant thereof is truncated at the 3′ untranslated region (UTR) of the PAH sequence or the variant thereof.7. The viral vector of claim 1 , wherein the therapeutic cargo portion further comprises a beta globin intron.8. The viral vector of claim 2 , wherein the therapeutic cargo portion further comprises at least one hepatocyte nuclear factor binding site.9. The viral vector of claim 8 , wherein the at least one hepatocyte nuclear factor binding site is disposed upstream of the prothrombin enhancer or downstream of the prothrombin enhancer.10. (canceled)11. The viral vector of claim 1 , wherein the PAH sequence or the variant thereof comprises a sequence having at least 80% identity with SEQ ID NO: 1; SEQ ID NO: 2; SEQ ID NO: 3; or SEQ ID NO: 4.12. (canceled)13. The viral vector of claim 2 , wherein the prothrombin enhancer comprises a sequence having at least 80% identity with SEQ ID NO: 5.14. (canceled)15. The viral vector of claim ...

LIPID NANOPARTICLE COMPOSITIONS AND METHODS FOR MRNA DELIVERY

Disclosed herein are compositions and methods for modulating the production of a protein in a target cell. The compositions and methods disclosed herein are capable of ameliorating diseases associated with protein or enzyme deficiencies. 140-. (canceled)41. A method for pulmonary delivery of messenger RNA (mRNA) for in vivo production of a protein in the lung comprising administering via inhalation from a nebulizer to the lung of a subject , a composition comprising an mRNA that encodes the protein ,wherein the mRNA is encapsulated within a lipid nanoparticle comprising one or more PEG-modified lipids, andwherein the administering of the composition results in detectable levels of the protein at least 72 hours after administration in the subject's lung.42. The method of claim 41 , wherein the mRNA comprises a 5′ untranslated region.43. The method of claim 42 , wherein the mRNA comprises a 3′ untranslated region.44. The method of claim 43 , wherein the mRNA comprises a cap structure.45. The method of claim 44 , wherein the mRNA comprises a poly A tail.46. The method of claim 45 , wherein the lipid nanoparticle comprises one or more cationic lipids.47. The method of claim 46 , wherein the lipid nanoparticle comprises one or more non-cationic lipids.48. The method of claim 47 , wherein the mRNA is unmodified.49. The method of claim 48 , wherein the composition is a reconstituted lyophilized composition.50. The method of claim 47 , wherein the modification comprises a modified nucleotide.51. The method of claim 50 , wherein the modified nucleotide is pseudouridine.52. The method of claim 51 , wherein the composition is a reconstituted lyophilized composition.53. The method of claim 41 , wherein the protein encoded by the mRNA is a polypeptide.54. The method of claim 41 , wherein the protein encoded by the mRNA is a peptide.55. The method of claim 48 , wherein the lipid nanoparticle has a size of less than about 100 nm.56. The method of claim 51 , wherein the lipid ...

Alpha-1-Antitrypsin (A1AT) Fusion Proteins and Uses Thereof

The invention relates to MIC-1 fusion proteins. More specifically it relates to compounds comprising fusion proteins comprising a MIC-1 protein or an analogue thereof at the C-terminus of the fusion protein and a functional variant of human A1AT (A1AT) at the N-terminus of the fusion protein connected via a peptide linker. The compounds of the invention have MIC-1 activity. The invention also relates to pharmaceutical compositions comprising such compounds and pharmaceutically acceptable excipients, as well as the medical use of the compounds. 1. A compound comprising a fusion protein of formula (I):{'br': None, 'A-B-C\u2003\u2003(I),'}wherein A is human A1AT or a functional variant thereof;wherein B is a peptide linker that is 10 to 100 amino acids in length;wherein C is a MIC-1 protein or an analogue thereof;wherein the C-terminus of the A1AT or a functional variant thereof is fused to the N-terminus of the peptide linker, andwherein the C-terminus of the peptide linker is fused to the N-terminus of the MIC-1 protein or analogue thereof.2. The compound according to claim 1 , wherein the peptide linker comprises the amino acid sequence [X-Y] claim 1 , wherein X is Asp or Glu; Y is Ala; m is from 2 to 4; and n is at least 5.3. The compound according to claim 2 , wherein X is Glu.4. The compound according to claim 2 , wherein m is 2-3 and n is 5-12.5. The compound according to claim 2 , wherein n is 7-12.6. The compound according to claim 2 , wherein the peptide linker comprises 1-4 Pro residues.7. The compound according to claim 6 , wherein the 1-4 Pro residues are in a midsection of the linker covering from approximately 20%-30% of the linker length.8. The compound according to claim 6 , wherein the peptide linker comprises 2 Pro residues.9. The compound according to claim 8 , wherein the peptide linker comprises 1 Pro residue.10. The compound according to claim 1 , wherein C is an analogue of MIC-1 displaying at least 85% sequence identity to native MIC-1 (SEQ ID ...

METHODS AND COMPOSITIONS FOR SINGLE CHAIN VARIABLE REGION ENOX2 ANTIBODIES FOR CANCER DETECTION AND DIAGNOSIS

Cancers of different cellular or tissue origins express different ENOX2 cancer isoforms or combinations of isoforms and shed these proteins into the circulation. Herein are disclosed methods both for cancer detection and diagnosis of particular origin, based on the patterns and molecular weights of the isoforms which allow the identification of the cell type and or tissue of origin of the neoplasm. Relative ENOX2 amounts are proportional to tumor burden and provide a reliable measure of response to therapy and disease progression. Also provided is the amino acid sequence to which the scFv antibodies bind as the molecular basis for the specificity of the test. 1. A cell that expressed an antibody that is a single chain variable region ENOX2-specific antibody fusion protein , wherein the antibody is fused with an alkaline phosphatase or other protein for single-step detection or imaging.2. The cell of claim 1 , wherein the cell is a bacterium.3. The cell of claim 1 , wherein the cell is a mammalian cell.4. A method for detecting cancer comprising:obtaining a sample from a patient suspected of comprising ENOX2 variants associated with a cancer; anddetecting isoforms of the ENOX2 protein using a single chain variable region ENOX2 antibody fusion protein, wherein the antibody is fused with an alkaline phosphatase or other single-step protein for detection or imaging, wherein expression of ENOX2 variants associated with cancer are indicative that the subject has or will develop that cancer.5. A method for producing a single chain variable region ENOX2 antibody comprising introducing into a cell a DNA sequence of SEQ ID NOS:1 and 2 claim 1 , and growing the cells to produce a composition containing a single chain variable region ENOX2 antibody.6. The method of claim 5 , wherein the DNA sequence includes DNA specifically producing the polypeptide linker sequence substantially as shown in SEQ ID NO:5 or 16.7. The method of claim 5 , wherein the DNA sequence comprises the ...

LIPID NANOPARTICLE COMPOSITIONS AND METHODS FOR MRNA DELIVERY

Disclosed herein are compositions and methods for modulating the production of a protein in a target cell. The compositions and methods disclosed herein are capable of ameliorating diseases associated with protein or enzyme deficiencies. 140-. (canceled)41. A method for delivery of messenger RNA (mRNA) for in vivo production of a factor IX (FIX) protein , comprising administering , to a human , a composition comprising an mRNA that encodes the FIX protein wherein the mRNA is encapsulated within a lipid nanoparticle , wherein the administering of the composition results in expression of the FIX protein encoded by the mRNA that is detectable in serum at least 72 hours after administration , and wherein the lipid nanoparticle comprises one or more PEG-modified lipids.42. The method of claim 41 , wherein the mRNA comprises a 5′ untranslated region.43. The method of claim 41 , wherein the mRNA comprises a 3′ untranslated region.44. The method of claim 43 , wherein the mRNA comprises a cap structure.45. The method of claim 44 , wherein the mRNA comprises a poly A tail.46. The method of claim 45 , wherein the lipid nanoparticle comprises one or more cationic lipids.47. The method of claim 46 , wherein the lipid nanoparticle comprises one or more non-cationic lipids.48. The method of claim 47 , wherein the mRNA is unmodified.49. The method of claim 48 , wherein the composition is a reconstituted lyophilized composition.50. The method of claim 47 , wherein the modification comprises a modified nucleotide.51. The method of claim 50 , wherein the modified nucleotide is pseudouridine.52. The method of claim 51 , wherein the composition is a reconstituted lyophilized composition.53. The method of claim 48 , wherein the composition is administered by intravenous injection.54. The method of claim 51 , wherein the composition is administered by intravenous injection.55. The method of claim 48 , wherein the composition is administered by intramuscular injection.56. The method of ...

BINDING FUSION PROTEINS, BINDING FUSION PROTEIN-DRUG CONJUGATES, XTEN-DRUG CONJUGATES AND METHODS OF MAKING AND USING SAME

The present invention relates to binding fusion protein compositions comprising targeting moieties linked to extended recombinant polypeptide (XTEN), binding fusion protein-drug conjugate compositions, and XTEN-drug conjugate compositions, isolated nucleic acids encoding the compositions and vectors and host cells containing the same, and methods of using such compositions in treatment of diseases, disorders, and conditions. 128-. (canceled)29. A binding fusion protein comprising:a first targeting moiety comprising a single-chain variable fragment (scFv) with specific binding affinity to CD3; anda first extended recombinant polypeptide (XTEN), wherein the first XTEN comprises a motif selected from the group consisting of SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, and SEQ ID NO: 9.30. The binding fusion protein of claim 29 , wherein the first XTEN comprises an amino acid sequence which has at least 90% sequence identity to a sequence selected from the group consisting of AE144 (SEQ ID NO: 41) claim 29 , AE288 (SEQ ID NO: 47) claim 29 , AE576 (SEQ ID NO: 54) and AE864 (SEQ ID NO: 59).31. The binding fusion protein of claim 30 , wherein the first XTEN comprises an amino acid sequence selected from the group consisting of AE144 (SEQ ID NO: 41) claim 30 , AE288 (SEQ ID NO: 47) claim 30 , AE576 (SEQ ID NO: 54) and AE864 (SEQ ID NO: 59).32. The binding fusion protein of claim 29 , wherein the first targeting moiety has specific binding affinity for an epsilon subunit of CD3 (CD3c).33. The binding fusion protein of claim 29 , wherein the first XTEN is positioned N-terminal of the first targeting moiety.34. The binding fusion protein of claim 29 , wherein the first XTEN is positioned C-terminal of the first targeting moiety.35. The binding fusion protein of claim 29 , further comprising a second targeting moiety that differs from the first targeting moiety claim 29 , wherein the second targeting moiety comprises a second single-chain variable fragment (scFv) with specific ...

Raav-based compositions and methods

The invention relates to isolated nucleic acids and rAAV-based compositions, methods and kits useful for treating genetic diseases (e.g., alpha-1 antitrypsin deficiency).

PROTEINS WITH MODIFIED GLYCOSYLATION AND METHODS OF PRODUCTION THEREOF

In one aspect, the disclosure provides proteins with modified glycosylation and methods of their production. In aspect, the disclosure provides transgenic animals and cells for the production of proteins with modified glycosylation. In some embodiment, the modified glycosylation is increased sialylation. 1. A method of producing protein with modified glycosylation , the method comprising:providing a transgenic mammal that has been modified to have increased expression of sialyl transferase in the mammary gland,wherein the transgenic mammal expresses a protein in the mammary gland, andharvesting the protein from the mammary gland of the transgenic mammal.2. A method of producing protein with modified glycosylation , the method comprising:providing a mammary epithelial cell that has been modified to have increased expression of sialyl transferase,wherein the mammary epithelial cell expresses a protein, andharvesting the protein from the mammary epithelial cells.3. The method of or , wherein the glycosylation is modified compared to a protein produced in a transgenic mammal or mammary epithelial cell that has not been modified to have increased expression of sialyl transferase.43. The method of any one of - claims 1 , wherein the modified glycosylation is increased sialylation.5. A method of producing protein with increased sialylation claims 1 , the method comprising:providing a transgenic mammal that has been modified to have increased expression of sialyl transferase in the mammary gland,wherein the transgenic mammal expresses a protein in the mammary gland, andharvesting the protein with increased sialylation from the mammary gland of the transgenic mammal.6. The method of or claims 1 , wherein the sialylation is increased compared to a protein produced in a transgenic mammal or mammary epithelial cell that has not been modified to have increased expression of sialyl transferase.76. The method of any one of - claims 1 , wherein the protein with modified ...

RECOMBINANT MUTANT (alpha)1-ANTITRYPSIN AND PREPARATION AND USES THEREOF

Provided is a new AAT triple-mutant, and methods to produce and purify the new entity. The new mutant is produced by a structure-based protein design to provide a more thermostable and oxidation-resistant agent for various pharmaceutical applications. The present invention also provides methods for expression, inclusion body refolding, and purification of the triple-mutant. Furthermore, the invention also provides methods for chemically modifying the purified drug candidate to provide a longer in vivo half-life and achieve better drug efficacy. 1. A recombinant mutant al-antitrypsin (AAT) polypeptide , wherein the polypeptide is a triple mutant F51L/M351V/M358V AAT polypeptide and/or a chemically modified form of α1-antitrypsin (AAT) polypeptide , having a chemical modification on cysteine at position 232 (Cys232) of wild-type or mutant AAT polypeptide , wherein the chemical modification on Cys232 comprises PEGylation or a fatty acid modification; wherein the fatty acid modification comprises Palmitoylation.2. The recombinant mutant AAT polypeptide of claim 1 , wherein the recombinant mutant AAT polypeptide comprises an amino acid sequence of SEQ ID NO:1 claim 1 , or an amino acid sequence with a deletion of one or more N-terminal amino acid residues of SEQ ID NO:1.3. The recombinant mutant AAT polypeptide of claim 3 , wherein the recombinant mutant AAT polypeptide comprises an amino acid sequence with a deletion of N-terminal amino acid residues at positions 1-5 of SEQ ID NO:1 claim 3 , or with a deletion of N-terminal amino acid residues at positions 1-10 of SEQ ID NO:1.4. The recombinant mutant AAT polypeptide of claim 1 , wherein the chemically modified form comprises a chemical modification at a specific site of the wild-type or F51L/M351V/M358V triple mutant.5. The recombinant mutant AAT polypeptide of claim 4 , wherein the chemical modification is performed at Cys232 or an N-terminal site.6. The recombinant mutant AAT polypeptide of claim 4 , wherein the ...

RAAV-BASED COMPOSITIONS AND METHODS FOR TREATING ALPHA-1 ANTI-TRYPSIN DEFICIENCIES

The invention relates to isolated nucleic acids and rAAV-based compositions, methods and kits useful for treating genetic diseases (e.g., alpha-1 antitrypsin deficiency). 1. A recombinant adenoviral associated vector (rAAV vector) comprising:(a) a first region that encodes one or more first miRNAs comprising a nucleic acid having sufficient sequence complementary with an endogenous mRNA of a subject to hybridize with and inhibit expression of the endogenous mRNA, wherein the endogenous mRNA encodes a first protein having a dominant negative or gain of function mutation; and(b) a second region encoding an exogenous mRNA that encodes a second protein, wherein the second protein has an amino acid sequence that is at least 85% identical to the first protein and does not have the dominant negative or gain of function mutation,wherein the one or more first miRNAs do not comprise a nucleic acid having sufficient sequence complementary to hybridize with and inhibit expression of the exogenous mRNA, and wherein the first region is positioned within between a portion of the second region encoding the last codon of the exogenous mRNA and a polyadenylation sequence of the exogenous mRNA.24-. (canceled)5. The rAAV vector of claim 1 , further comprising a third region encoding a one or more second miRNAs comprising a nucleic acid having sufficient sequence complementary to hybridize with and inhibit expression of the endogenous mRNA claim 1 , wherein the third region is positioned within an untranslated portion of the second region.67-. (canceled)8. The rAAV vector of claim 5 , wherein the third region is between the first codon of the exogenous mRNA and a position 1000 nucleotides upstream of the first codon.9. The rAAV vector of claim 1 , wherein the first region encodes two first miRNAs.10. The rAAV vector of claim 1 , wherein the first region encodes three first miRNAs.11. The rAAV vector of claim 5 , wherein the third region encodes two second miRNAs.12. The rAAV vector of ...

NOVEL GROWTH HORMONE RECEPTOR ANTAGONIST AND FUSION PROTEIN THEREOF

The present invention relates to a growth hormone receptor antagonist comprising a growth hormone variant which is modified by substitution of one or more amino acids of growth hormone. Further, the growth hormone receptor antagonist of the present invention may further comprise a long-acting carrier which is fused to the growth hormone variant. The growth hormone receptor antagonist may have strong binding potency to growth hormone receptor and may exhibit a long-lasting antagonistic action. 1. A growth hormone receptor antagonist comprising a growth hormone variant in which one or more amino acids in an amino acid sequence of growth hormone are substituted with another amino acid.2. The growth hormone receptor antagonist of claim 1 , wherein the substitution with another amino acid comprises substitution of the 46amino acid from the N-terminus with lysine claim 1 , substitution of the 120amino acid from the N-terminus with lysine or arginine claim 1 , or a combination thereof.3. The growth hormone receptor antagonist of claim 2 , wherein the substitution with another amino acid further comprises substitution at any one or more positions selected from the group consisting of the 18amino acid claim 2 , the 21amino acid claim 2 , the 54amino acid claim 2 , the 64amino acid claim 2 , the 167amino acid claim 2 , the 168amino acid claim 2 , the 171amino acid claim 2 , the 172amino acid claim 2 , the 174amino acid claim 2 , the 176amino acid claim 2 , and the 179amino acid from the N-terminus.4. The growth hormone receptor antagonist of claim 2 , wherein the substitution with another amino acid further comprises substitution of the 174amino acid from the N-terminus with serine claim 2 , substitution of the 21amino acid from the N-terminus with asparagine claim 2 , or a combination thereof.5. The growth hormone receptor antagonist of claim 1 , wherein the substitution with another amino acid comprises any one or more substitutions selected from the group consisting of ...

LIPID NANOPARTICLE COMPOSITIONS AND METHODS FOR MRNA DELIVERY

Disclosed herein are compositions and methods for modulatimi the production of a protein in a target cell, The compositions and methods disclosed herein are capable of ameliorating diseases associated with protein or enzyme deficiencies.