ХИМИЧЕСКИЕ СОЕДИНЕНИЯ

Изобретение относится к области медицины, в частности онкологии. Сущность изобретения составляют улучшенные системы направленной терапии опухолей с использованием фермента и пролекарственного соединения, а в частности антитело-опосредованной терапии с использованием фермента и пролекарственного соединения (ADEPT). В системе используемый фермент представляет собой мутированную форму фермента клетки-хозяина, где натуральный фермент клетки-хозяина, такой как рибонуклеаза, располагает свой натуральный субстрат посредством взаимодействия пары ионов. При этом в данной системе мутированного фермента и комплементарного пролекарственного соединения это взаимодействие является обратным (т.е. обратной полярности). Техническим результатом изобретения является расширение арсенала противоопухолевых средств. 9 с. и 11 з.п. ф-лы, 33 ил.

ТРИПЕПТИДИЛПЕПТИДАЗЫ, СПОСОБНЫЕ ВОЗДЕЙСТВОВАТЬ НА СВЯЗИ С УЧАСТИЕМ ПРОЛИНА, И ИХ ПРИМЕНЕНИЯ

Изобретение относится к области биотехнологии. Описан способ получения гидролизата. Заявленный способ включает стадию смешивания белка или его части с эндопротеазой и толерантной к пролину трипептидилпептидазой, обладающей экзопептидазной активностью, где указанная толерантная к пролину трипептидилпептидаза способна к отщеплению трипептидов с N-конца пептидов, имеющих пролин в P1' или аминокислоту, выбранную из аланина, аргинина, аспарагина, аспарагиновой кислоты, цистеина, глутамина, глутаминовой кислоты, глицина, гистидина, изолейцина, лейцина, лизина, метионина, фенилаланина, серина, треонина, триптофана, тирозина, валина или синтетических аминокислот в положении P1'. Изобретение расширяет арсенал средств для получения гидролизата. 14 з.п. ф-лы, 28 ил., 13 табл., 24 пр.

ГЕН НОВОЙ СЕРИНОВОЙ ПРОТЕАЗЫ, РОДСТВЕННОЙ DPPIV

Изобретение относится к области биотехнологии и может быть использовано в фармакологии и медицине. Клонирована и определена последовательность ДНК, кодирующая новый вид сериновой протеазы человека (DPRP-2), родственной дипептидилолигопептидазе IV, гомология между которыми на уровне выведенной аминокислотной последовательности определена как 39%. С помощью технологии рекомбинантных ДНК получена рекомбинантная форма DPRP-2 и исследованы кинетические свойства нового фермента (субстратная специфичность, значения Km, ингибиторы активности и т.п.). Осуществление изобретения обеспечивает новые возможности при поиске и создании новых терапевтических средств для лечения широкого круга заболеваний человека. 2 н. и 3 з.п. ф-лы, 4 ил., 6 табл.

СПОСОБ ПОЛУЧЕНИЯ ДИПЕПТИДА (ВАРИАНТЫ)

... 1. Способ получения дипептида, включающий обеспечение воздействия белка, обладающего пролиниминопептидазной активностью, на эфир L-аминокислоты и L-аминокислоту с образованием дипептида, где белок, обладающий пролиниминопептидазной активностью, происходит из микроорганизма, принадлежащего к роду Pseudomonas. 2. Способ по п.1, где белок, обладающий пролиниминопептидазной активностью, происходит из Pseudomonas putida. 3. Способ по п.1 или 2, где эфир L-аминокислоты представляет собой один или несколько представителей, выбранных из группы, состоящей из эфира L-аланина, эфира глицина, эфира L-валина, эфира L-изолейцина, эфира L-метионина, эфира L-фенилаланина, эфира L-серина, эфира L-треонина, эфира L-глутамина, эфира L-тирозина, эфира L-аргинина, α-эфира L-аспарагиновой кислоты, β-эфира L-аспарагиновой кислоты, эфира L-лейцина, эфира L-аспарагина, эфира L-лизина, α,β-диметилового эфира L-аспарагиновой кислоты, и γ-эфира L-глутамина. 4. Способ по п.1 или 2, где L-аминокислота представляет собой ...

Способ количественного определения амилолитической активности ферментов по гидролизу субстрата, иммобилизованного в полиакриламидном геле

ФАРМАЦЕВТИЧЕСКИЕ ПРИМЕНЕНИЯ ФЕРМЕНТНОЙ КОМПОЗИЦИИ, ВЫДЕЛЕННОЙ ИЗ АНТАРКТИЧЕСКОГО КРИЛЯ

Неиммуногенные ферментные композиции, выделенные из антарктического криля и проявляющие как эндо-, так и экзопептидазную активность, применимы для приготовления лекарственных препаратов и фармацевтических композиций для лечения большого разнообразия заболеваний человека и животных, таких как инфекции, воспаления, раковые заболевания, ВИЧ/СПИД, боль, полипы, бородавки, геморрои, бляшки, морщины, утончение волос, аллергический зуд, спайкообразование, глазные болезни, такие как катаракта, глаукома, и т.д.

СПОСОБ ЭКСПРЕССИИ

... 1. Способ получения белка посредством микроорганизма, содержащий стадии:(a) введение в микроорганизм первой экспрессионной конструкции, которая кодирует белок;(b) введение в микроорганизм второй экспрессионной конструкции, которая кодирует вспомогательную протеазу, которая отличается от белка и которая содержит аминокислотную последовательность, которая по меньшей мере на 50% идентична аминокислотной последовательности, обозначенной как SEQ ID NO.1, где вспомогательная протеаза имеет протеолитическую активность;(c) экспрессию белка и вспомогательной протеазы в микроорганизме.2. Способ по п.1, отличающийся тем, что белок естественным образом не присутствует в микроорганизме, и/или вспомогательная протеаза естественным образом не присутствует в микроорганизме.3. Способ по п.1, отличающийся тем, что вспомогательная протеаза замещает по меньшей мере одну кодируемую хромосомой протеазу в микроорганизме, или где вспомогательная протеаза экспрессируется в микроорганизме в дополнение к кодируемым ...

Verfahren zur effizienten Herstellung von 7-ADCA via 2-(carboxyethylthio)acetyl-7-ADCA und 3-(carboxymethylthio)propionyl-7-ADCA

Carboxypeptidase-ähnliches Protein aus Knochen und Verfahren zur dessen Herstellung

INHIBITOREN VON KATALYTISCHEN ANTIKÖRPERN

Methode zur Herstellung von menschlichen Wachstumshormonen

Rekombinante Carboxypeptidase B und ihre Aufreinigung

ACID CARBOXYPEPTIDASE AND THE PREPARATION THEREOF

... 1,261,363. Acid carboxypeptidase. KIKKOMAN SHOYU CO. Ltd. 15 April, 1969 [9 Dec., 1968], No. 19134/69. Heading C3H. An acid carboxypeptidase reacts hydrolytically with the terminal peptide linkage of a protein or peptide adjacent to the terminal free carboxyl group to liberate the terminal amino acid as free amino acid at a pH of from 1À5 to 5À5, contains no metal, has a molecular weight of about 122,000 and is inactive at 60‹ C. or more or a pH of 7 or more. It may be prepared from Aspergillus usamii, A. saitoi, A. riger, A. inuii, A. aureus, A. awamori, A. nakazawaii, A. oryzae or A. sojae by aerobic culture: the enzyme occurs in the culture medium and can be worked up by conventional methods.

Engineered E2

Treatment of diseases related to hyperactivity of the complement system

LYS-AMINOPEPTIDASE PEPN FROM LACTOBACILLUS DELBRUKII SSP.LACTIS,NUCLEIC ACIDS CODING FOR IT,AND ITS USE IN FERMENTATION PROCESSES

METHOD OF PURIFYING A PROTEIN

Combination of complement factors i and h, and vector encoding thereof

Process for the isolation of crystalline imipenem.

The present invention relates to a cost effective and industrially advantageous process for the preparation of imipenem of high purity. The process for the isolation of pure crystalline imipenem monohydrate of Formula I, comprises contacting crude imipenem with a water miscible organic solvent, either alone or in admixture with water and recovering pure crystalline imipenem monohydrate.

Process for the isolation of crystalline imipenem.

Process for the preparation of amorphouse cilastatin sodium

The present invention relates to a cost effective and industrially advantageous process for the preparation of amorphous cilastatin sodium.

TOBACCO PROTEASE GENES

Novel carboxypeptidase of cocoa.

Process for the preparation of amorphous cilastatin sodium.

Process for the isolation of crystalline imipenem.

Process for the preparation of amorphouse cilastatin sodium.

Process for the isolation of crystalline imipenem.

TOBACCO PROTEASE GENES

Process for the preparation of amorphouse cilastatin sodium.

REKOMBINANTE CARBOXYPEPTIDASE B AND THEIR AUFREINIGUNG

WITHOUT PRESSURE TAKING PLACE PRETREATMENT, ENZYMATIC HYDROLYSIS AND FERMENTATION OF WASTE PARLIAMENTARY GROUPS

ACID-ACTIVE FUNGALE PROTEASE

RECUPERATION OF A PROTEIN AT HIGH PH VALUE

17867, A HUMAN AMINOPEPTIDASE

ACE2-AKTIVIERUNG FOR THE TREATMENT OF HEART, LUNGS, AND KIDNEY ILLNESS AND HYPERTENSION

NEW MUSHROOM PROTEINS AND THESE CODING NUCLEIC ACIDS

NEW BLUTGERINNSELAUFLÖSENDE PROTEINS AND YOUR PRODUCTION FROM THE BLUTEGEL HIRUDO MEDICINALIS.

FOR HUMAN ANGIOTENSION UMFORMUNGSENZYM (ACE) CODING NUCLEIC ACID AND YOUR APPLICATIONS, IN PARTICULAR TO IN-VITRO-DIAGNOSIS OF HYPERTENSION.

DIPEPTIDASE, THEIR ISOLATION FROM LACTIC ACID BACTERIA, IMMUNE BODIES AGAINST THIS DIPEPTIDASE, USE THESE DIPEPTIDASE AND THIS IMMUNE BODIES.

CARBOXYPEPTIDASEN OUT ASPERGILLUS ORYZAE AND THESE CODING NUCLEIC ACIDS

POLYPEPTIDE WITH AMINOPEPTIDASEAKTIVITÄT AND FOR THESE CODING NUCLEIC ACIDS

PROCEDURE FOR THE DETERMINATION THE KARDIOVASCULÄREN OF THE STATUS AND COMPOSITIONS FOR THEIR USE

GRANULATES CONTAINING HYDRATISIERTES CHECK MATERIAL

NEW AMINOPEPTIDASE FROM STREPTOMYCES

CHEMICAL MEANS

METHOD FOR THE PRODUCTION OF HUMAN GROWTH HORMONES

PRODUCTION THAT AMINOPEPTIDASEN OUT ASPERGILLUS THE NIGER

ENZYME WITH AMINOPEPTIDASEACTIVITÄT

CARBOXYPEPTIDASE SIMILAR PROTEIN FROM BONES AND PROCEDURES FOR ITS PRODUCTION

Systems and methods for producing collagen 7 compositions

The present disclosure provides production systems and host cells to produce collagen 7 compositions comprising recombinant collagen 7 and/or functional variants thereof. The host cells are genetically engineered to stably express rCol7 and functional variants thereof. The collagen 7 composition can be used to restore collagen 7 levels in a subject in need, and for preventing, preventing the progression of, alleviating, and delaying the on-set of a skin condition, e.g., skin wound associated with dystrophic epidermolysis bullosa (DEB).

Complement factor I and complement factor I cofactor, vectors encoding therefor and therapeutic use

A product comprising (i) a Complement Factor I (CFI) cofactor; and (ii) Complement Factor I (CFI), or nucleotide sequences encoding therefor, as a combined preparation for simultaneous, separate or sequential use in therapy.

Treatment of fibroses and liver disorders

Novel serine protease genes related to DPPIV

Mirac proteins

This disclosure relates to a method of generating conditionally active biologic proteins from wild type proteins, in particular therapeutic proteins, which are reversibly or irreversibly inactivated at the wild type normal physiological conditions. For example, evolved proteins are virtually inactive at body temperature, but are active at lower temperatures.

Modulation of cop9 signalsome isopeptidase activity

Methods for treating endocrine disorders

Enzyme and snp marker for disease

Methods and apparatuses for purification

NOVEL PURIFIED POLYPEPTIDES INVOLVED IN PROTEIN SYNTHESIS AND MODIFICATION

NOVEL ENZYME FORMING PEPTIDE, MICROORGANISM PRODUCING THE SAME AND PROCESS FOR PRODUCING DIPEPTIDE USING THEM

Fusion proteins and methods for stimulating plant growth, protecting plants, and immobilizing Bacillus spores on plants

The present invention is generally directed to fusion proteins containing a targeting sequence that targets the fusion protein to the exosporium of a Bacillus cereus family member. The invention also relates to recombinant Bacillus cereus family members expressing such fusion proteins and formulations containing the recombinant Bacillus cereus family members expressing the fusion proteins. Methods for stimulating plant growth, for protecting plants from pathogens, and for enhancing stress resistance in a plant by applying the recombinant Bacillus cereus family members or the formulations to plants or a plant growth medium are also described. The invention also relates to methods for immobilizing spores of a recombinant Bacillus cereus family member expressing a fusion protein on plants.

Multiple proteases deficient filamentous fungal cells and methods of use thereof

The present disclosure relates to compositions and methods useful for the production of heterologous proteins in filamentous fungal cells.

Use of proline tolerant tripeptidyl peptidases in feed additive compositions

A method of preparing a feed additive composition comprising: (a) admixing at least one proline tolerant tripeptidyl peptidase predominantly having exopeptidase activity wherein said proline tolerant tripeptidyl peptidase is capable of cleaving tri-peptides from the N-terminus of peptides having (i) (A)Proline at P1; and (B) An amino acid selected from alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, serine, threonine, tryptophan, tyrosine, valine or synthetic amino acids at P1; or (ii) (a') Proline at PV; and (b') An amino acid selected from alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, serine, threonine, tryptophan, tyrosine, valine or synthetic amino acids or amines at PV; and one or more ingredients selected from the group consisting of a salt, polyol including sorbitol ...

Novel polypeptides with improved proteolytic stability, and methods of preparing and using same

The present invention includes methods of improving proteolytic stability of a polypeptide, comprising alkylating at least one selected from the group consisting of a N-terminus amino group, the NH group of the N-terminus first internal amide bond, another primary amino group, a thiol group and a thioether group within the polypeptide. The present invention further includes polypeptides incorporating such chemical modifications.

Catalytic antibody components

Selection methods for genetically-modified T cells

In some aspects, isolated transgenic cells (e.g., transgenic T cells) are provided that comprise or express a transgene and DHFRFS and/or TYMSss. Methods for selecting transgeneic cells are also provided.

Cloning of an insulin-dependent membrane aminopeptidase from glut-4 vesicles

Method for extra-cellular expression of protein

Recovery of a protein at high ph

Regulation of human carboxypeptidase-like enzyme

Modified kluyveromyces yeasts, their preparation and use

Compositions and methods for interfering with hepatitis b virus infection

Carboxypeptidases from aspergillus oryzae and nucleic acids encoding same

ISOLATION, PURIFICATION, AND CHARACTERIZATION OF THE AMINOPEPTIDASES: AP2, AP1, AND APX

22196, A NOVEL HUMAN AMINOPEPTIDASE

The present invention relates to a newly identified human aminopeptidase. The invention also relates to polynucleotides encoding the aminopeptidase. The invention further relates to methods using the aminopeptidase polypeptides and polynucleotides as a target for diagnosis and treatment in aminopeptidase- related disorders. The invention further relates to drug-screening methods using the aminopeptidase polypeptides and polynucleotides to identify agonists and antagonists for diagnosis and treatment. The invention further encompasses agonists and antagonists based on the aminopeptidase polypeptides and polynucleotides. The invention further relates to procedures for producing the aminopeptidase polypeptides and polynucleotides.

PROCESS FOR THE PREPARATION OF AMORPHOUS CILASTATIN SODIUM

The present invention relates to a cost effective and industrially advantageous process for the preparation of amorphous cilastatin sodium.

DIPEPTIDYL PEPTIDASES

A novel human dipeptidyl aminopeptidase (DPP8) with structural and functional similarities to DPPIV and fibroblast activation protein (FAP) has been identified and characterized. The novel peptide comprises the sequence ID NO: 1, the amino acid sequences His736GlyTrpSerTyrGlyGlyTyrLeu, Leu816AspGluAsnValHisPheAlaHis, Glu847ArgHisSerIleArg, Phe225ValLeuGlnGluGluPhe, or a sequence having at least 60 % homology with, and the substrate specificity of SEQ ID NO:1. A gene encoding the dipeptidyl peptidase and uses of the dipeptidyl peptidase.

USE OF COMPLEMENT FACTOR I FOR THE TREATMENT OF DISEASES RELATED TO HYPERACTIVITY OF THE COMPLEMENT SYSTEM

Raising the level of Factor I above physiological levels can be used to treat diseases in which the underlying pathology is linked to overactivity of the C3b-feedback cycle and the generation and pro-inflammatory effects of iC3b. Methods, agents, and compositions for treatment of such diseases are described.

HIGH-THROUGHPUT ASSAY FOR EVALUATING DIPEPTIDYL PEPTIDASE I ACTIVITY

A method of screening a compound which modulates dipeptidyl peptidase I (DPPI) activities comprises the steps of adding a peptide substrate of DPPI to a reaction mixture which comprises DPPI and a compound, wherein the peptide substrate of DPPI has at least 3 amino acids and binds to a binding site of DPPI in addition to the S1-S2 site; and measuring the molecular weight of the substrate, wherein a change in the molecular weight of the substrate is indicative of the presence of DPPI activity.

METHODS AND COMPOSITIONS FOR RESISTANCE TO CYST NEMATODES IN PLANTS

The disclosure relates to methods and compositions for producing plants or plant cells that exhibit improved cyst nematode resistance.

ACID FUNGAL PROTEASES

NOVEL PEPTIDE-FORMING ENZYME GENE

ANTI-HUMAN TRANSFERRIN RECEPTOR ANTIBODY PERMEATING BLOOD-BRAIN BARRIER

Disclosed is a means that can be used for modifying a biologically or pharmacologically active substance from a form incapable of permeating the blood-brain barrier to a form capable of permeating the blood-brain barrier. Also disclosed is a modified substance obtained by using the means. The means is anti-human transferrin receptor antibody and the modified substance is a molecular conjugate of a biologically active protein or a pharmacologically active low-molecular-weight compound and anti-human transferrin receptor antibody.

SELECTION METHODS FOR GENETICALLY-MODIFIED T CELLS

In some aspects, isolated transgenic cells (e.g., transgenic T cells) are provided that comprise or express a transgene and DHFRFS and/or TYMSSS. Methods for selecting transgenic cells are also provided.

TREATMENT OF FIBROSES AND LIVER DISORDERS

The present invention relates to ACE2 for the therapeutic treatment or prevention of a fibrosis or liver disorder.

BMP-4 PEPTIDES & METHODS OF USE

The invention relates to truncated growth factors and variants thereof. The invention also relates to methods of making and using the truncated growth factors.

METHODS AND MEANS FOR STUDYING GENETIC POLYMORPHISM IN THE ANGIOTENSIN CONVERTING ENZYME I

... 2111953 9300360 PCTABS00161 L'invention concerne un procédé de détection du polymorphisme de l'enzyme de conversion (ECA) par une technique d'amplification de chaînes. Il a pour objet la détection de la présence ou de l'absence de la séquence nt.1451-1738 dans le gène ECA humain.

ENZYMATIC PROCESS FOR THE CONVERSION OF PREPROINSULINS INTO INSULINS

An enzymatic process for the conversion of preproinsulins into insulins A process for the specific hydrolysis of the amino-acid chain of preproinsulins to give the corresponding inter- mediates using clostripain is described, and these intermediates can, where appropriate, be converted with carboxypeptidase B into the corresponding insulins.

ENZYMATIC PROCESS FOR THE CONVERSION OF PREPROINSULINS INTO INSULINS

HOE 90/F 263 of the disclosure: An enzymatic process for the conversion of preproinsulins into insulins A process for the specific hydrolysis of the amino-acid chain of preproinsulins to give the corresponding intermediates using clostripain is described, and these intermediates can, where appropriate, be converted with carboxypeptidase B into the corresponding insulins.

DICTYOSTELIUM DIPEPTIDYLAMINOPEPTIDASE

The present invention is directed to a novel dipeptidylaminopeptidase isolated from the cellular slime mold, Dictyostelium discodeum. The novel DAP enzyme, dDAP, displays an activity which is somewhat similar to both DAPI and DAP-III but is highly distinctive from these enzymes in physical and other enzymatic characteristics. The invention is also directed to methods for using the dDAP enzyme to remove dipeptides from the N-terminus of recombinantly produced precursor proteins or peptides. In addition, the invention relates to methods for isolating and purifying the dDAP enzyme from cultures of D. discoedeum.

STRAIN OF SACCHAROMYCES CEREVISIAE PRODUCING AN HETEROLOGOUS PROTEIN AND PROCESS FOR THE PREPARATION OF SAID HETEROLOGOUS PROTEIN BY FERMENTATION OF SAID STRAIN

STRAIN OF SACCHAROMYCES CEREVISIAE PRODUCING AN HETEROLOGOUS PROTEIN AND PROCESS FOR THE PREPARATION OF SAID HETEROLOGOUS PROTEIN BY FERMENTATION OF SAID STRAIN

L'invention concerne une nouvelle souche de Saccharomyces cerevisiae productrice d'une protéine hétérologue, et plus particulièrement d'hirudine caractérisée par une suppression de la fonction protéolytique codée par le gène PRC1. Plus précisément, cette déficience est due à une mutation ou plus particulièrement à une délétion du gène de structure PRC1.

INHIBITORS OF CATALYTIC ANTIBODIES

Specific, selective inhibitors of catalytic antibodies both synthetic and naturally occurring, their use and compositions thereof are disclosed. In particular, an inhibitor preventing the hydrolysis of the peptide bond between amino acid residues 16 and 17 in the neurotransmitter vasoactive intestinal peptide (VIP) by an anti-VIP catalytic autoantibody is disclosed.

PROTEASES FROM STREPTOMYCES AND USE THEREOF IN PROTEIN EXPRESSION SYSTEMS

A family of proteases endogenous to Streptomyces cells degrades exogenous proteins secreted from Streptomyces host cells. The previously unidentified proteases include (1) tripeptidyl aminopeptidase designated "Tap", (2) tripeptidyl aminopeptidase designated "Ssp", (3) X-Pro-metalloendoproteinase designated "XP-Mep", and (4) other proteases derived from Streptomyces which degrade certain substrates under certain conditions. Degradation was alleviated by selective inhibition of secreted proteases or by using improved strains which lack or have impaired degradation proteases. An irreversible inhibitor was designed based upon the mechanism and substrate specificity of the target protease. Hosts expressing proteases were also produced. Uses of the proteases include immunoassays and proteolytic removal of peptides and polypeptides to improve secretion of exogenous proteins.

THERAPY

The present invention relates to improvements in targetted enzyme prodrug therapy including antibody-directed enzyme prodrug therapy (ADEPT), it particularly relates to novel enzymes and prodrugs for use in ADEPT.

EXPRESSION OF PROTECTIVE ANTIGENS

The invention provides a method for the expression of an enzyme antigen which in nature is parasite gut membrane bound enzyme or a fragment thereof having similar enzyme and/or antigenic activity, wherein mammalian host cells are transfected with a vector adapted to express said enzyme antigen or fragment thereof, characterised in that prior to transformation the host cells are substantially free from endogenous enzymes having (a) the same function and (b) the same cell membrane integration or lack of integration, as said parasite enzyme antigen or fragment thereof. Such a method is particularly useful for the expression of helminthic integral membrane aminopeptidase antigens.

A NOVEL FORM OF DIPEPTIDYLPEPTIDASE IV (CD26) FOUND IN HUMAN SERUM, ANTIBODIES THERETO, AND USES THEREFOR

A circulating, soluble form of DPPIV/CD26 isolated from human serum is disclosed. The serum form shares similar enzymatic and antigenic properties with the ubiquitous membrane form. However, in several biochemical aspects there are distinct differences. In particular, the circulating serum form has a molecular weight of 175 kDa (in contrast to the 105 kDa molecular weight of the membrane form), and it does not bind Adenosine Deaminase Type-1. Nevertheless, the circulating form expresses functional dipeptidylpeptidase IV activity and retains the ability to costimulate the T lymphocyte response to recall antigen. Circulating DPPIV has been determined to be the soluble form of a 175 kDa DPPIVCD26-related molecule rapidly expressed on the surface of activated T cells, prior to the expression of 105 kDa CD26. Although 105 kDa membrane type CD26 may be found in the serum in small amounts, the majority of serum DPPIV activity is provided by a novel peptidase structurally distinct from 105 kDa ...

BOVINE DIPEPTIDYLAMINOPEPTIDASE 1

The present invention provides the cDNA sequence encoding bovine dipeptidylaminopeptidase 1. The invention demonstrates that bovine DAP 1 coded by a single cDNA. The mature protein is derived from a single polypeptide consisting of a signal peptide, and a major polypeptide which is processed to generate the a subunit, b subnit and g subunit.

HYPERTENSION MARKERS

The present invention relates to a method of detecting for an individual mammal a high likelihood of having hypertension or a genetic predisposition for having hypertension, the method comprising determining the presence in a biological sample from the individual mammal of at least one marker protein which is indicative of a high likelihood of having hypertension or a genetic predisposition for having hypertension, wherein the marker protein is selected from the group consisting of the proteins shown in Table 1. In a further aspect the invention relates to novel proteins, nucleic acid fragments and use of said proteins or nucleic acid fragments for detection of whether an individual mammal has a high likelihood of having hypertension or a genetic predisposition for having hypertension as well as test kits for this purpose.

METHODS OF PRODUCING A RECOMBINANT PROTEIN

The subject invention relates to methods of processing and producing recombinant proteins in vivo, for example, recombinant phosphorylated betacasein. The subject invention also relates to the proteins produced by these methods as well as to uses of these proteins.

Pharmaceutical composition containing nuclear factor involved in proliferation and differentiation of central neuronal cells

With an aim to provide a novel factor inducing proliferation of neural stem cells and differentiation of these cells into nerve cells, a pharmaceutical composition comprising 1) CRBN, 2) a nucleic acid encoding CRBN, or 3) a stem cell or a neural progenitor cell in which CRBN is expressed, a method including administering the pharmaceutical composition to a non-human animal and inducing proliferation of neural stem cells or neural progenitor cells of the non-human animal and differentiation of these cells into nerve cells, and a method for screening for a therapeutic drug for a disease of cerebral cortex or a surgical injury of cerebral cortex, using CRBN, are provided.

Endoglucanase variants

The present invention relates to variant endoglucanases having improved thermoactivity, improved thermostability, and improved viscosity reduction activity over wild-type M. thermophila endoglucanase.

Accelerated fungal growth

The present invention relates to a process for the preparation of fermented food wherein a carboxypeptidase preparation is used to obtain accelerated fungal growth.

Collagen mixture and method of making the same

A collagen mixture having a portion of unhydrolyzed eggshell membrane collagen and Avian collagen.

Method for protein production in filamentous fungi

The present invention relates to a method for genetically modifying a filamentous fungus host for improved protein production. The method comprises that a filamentous fungus host is genetically modified to overexpress or to be deficient of specific genes. The invention relates also to the modified hosts. Furthermore, the invention relates to a method for improved production or for producing an improved composition of proteins, such as cellulases, hemicellulases, other proteins involved in the degradation of lignocellulosic material, or other proteins, in a filamentous fungus host. 1. A method to genetically modify a filamentous fungus host for improved protein production , said method comprising:{'i': Trichoderma, Aspergillus, Fusarium, Neurospora, Talaromyces, Phanerochaete, Chrysosporium', 'Penicillium, 'genetically modifying a filamentous fungus host to overexpress with increased amount or activity, or to be deficient with reduced or lacking amount or activity, of one or more genes selected from the group consisting of genes tre77513 (SEQ ID NO:1), tre80291 (SEQ ID NO:2), tre41573 (SEQ ID NO:3), tre74765 (SEQ ID NO:4), and tre64608 (SEQ ID NO:5); or of the closest homologue of at least one of said genes in or ; or of a fragment or derivative of any of said genes or other nucleotide sequence hybridizing under stringent conditions to at least one of said genes or said homologues, said host being capable of increased or decreased production of cellulase, hemicellulase, other proteins involved in degradation of lignocellulosic material and/or other proteins as compared to the parental strain.'}2Trichoderma, Aspergillus, Fusarium, Neurospora, Talaromyces, Phanerochaete, ChrysosporiumPenicillium. The method according to claim 1 , wherein the filamentous fungus host is genetically modified to overexpress one or more genes selected from the group consisting of genes tre77513 (SEQ ID NO:1) claim 1 , tre80291 (SEQ ID NO:2) claim 1 , tre41573 (SEQ ID NO:3) claim 1 , ...

Protein Fusion Constructs Possessing Thrombolytic and Anticoagulant Properties

The present invention discloses novel hybrid proteins that have both plasminogen activator and anti-thrombotic properties, including clot specific action, that renders these as highly advantageous for the treatment of circulatory disorders involving fibrin clot formation due to underlying tissue damage in the blood vessels leading to myocardial infarction, strokes etc. Also disclosed are new proteins, and methods of obtaining the same, that help to dissolve blood clots by activating plasminogen in a plasmin or thrombin dependent manner and also inhibit both the activity and generation of thrombin through the intrinsic pathway of blood coagulation. 1. A chimeric protein construct comprising the 4 , 5 , and 6 epidermal growth factor-like domains (EGF 4 ,5 ,6) of thrombomodulin fused to a thrombolytic protein selected from the group consisting of a streptokinase , a tissue plasminogen activator , a staphylokinase , a urokinase , and derivatives and analogs thereof.2. The chimeric protein construct according to claim 1 , where said thrombomodulin EGF 4 claim 1 ,5 claim 1 ,6 domains are fused to said thrombolytic protein claim 1 , or derivative or analog thereof at the N-terminus claim 1 , C-terminus claim 1 , or both N- and C-termini of said thrombolytic protein claim 1 , or derivative or analog thereof.3. The chimeric protein construct according to claim 1 , wherein said thrombolytic protein comprises a streptokinase with one or more amino acid substitutions claim 1 , insertions claim 1 , deletions claim 1 , or truncations claim 1 , and wherein the construct possesses plasminogen activation claim 1 , thrombin inhibition claim 1 , and anticoagulant protein C pathway activation activity.4. The chimeric protein construct according to claim 1 , where the thrombomodulin EGF 4 claim 1 ,5 claim 1 ,6 domains are fused between the alpha and beta or beta and gamma domains of said streptokinase or between the alpha and beta or beta and gamma domains of a streptokinase derivative ...

Protease Variants

The invention relates to a novel 3D structure encoding a as well as to variants of parent protease homologous to proteases, preferably of improved thermostability and/or with an amended temperature activity profile. The invention also relates to DNA sequences encoding such variants, their production in a recombinant host cell, as well as methods of using the variants, in particular within the field of animal feed and detergents. The invention furthermore relates to methods of generating and preparing protease variants of amended properties. 134-. (canceled)35. A variant of a parent protease , comprising a substitution in at least one position of at least one region selected from the group of regions consisting of:6-18; 22-28; 32-39; 42-58; 62-63; 66-76; 78-100; 103-106; 111-114; 118-131; 134-136; 139-141; 144-151; 155-156; 160-176; 179-181; and 184-188; (a) the variant has protease activity; and', '(b) each position corresponds to a position of amino acids 1 to 188 of SEQ ID NO: 2; and', '(c) the variant has a percentage of identity to amino acids 1 to 188 of SEQ ID NO: 2 of at least 60%., 'wherein'}36. An animal feed additive comprising at least one protease variant of claim 35 , and(a) at least one fat soluble vitamin;(b) at least one water soluble vitamin; and/or(c) at least one trace mineral.37. An animal feed composition having a crude protein content of 50 to 800 g/kg and comprising the protease variant of .38. A method for improving the nutritional value of an animal feed claim 35 , comprising adding a protease variant of to the feed.39. A method for the treatment of proteins claim 35 , comprising the step of adding the protease variant of to at least one protein or protein source. This application is a divisional of U.S. application Ser. No. 12/984,826 filed on Jan. 5, 2011, now allowed, which is a divisional of U.S. application Ser. No. 10/574,554 filed on Apr. 3, 2006, now U.S. Pat. No. 7,892,808, which is a 35 U.S.C. 371 national application of PCT/DK2004 ...

POLYPEPTIDE FOR INHIBITING METASTASIS, USES THEREOF, AND PHARMACEUTICAL COMPOSITIONS CONTAINING THE SAME

A polypeptide for inhibiting, treating or diagnosing metastasis and an use thereof are disclosed, wherein the polypeptide is a polypeptide, a derivative polypeptide, or a mutated polypeptide of a fibronectin-binding domain of dipeptidyl peptidase IV. In addition, a pharmaceutical composition treating or diagnosing metastasis is also disclosed, which comprises the aforementioned polypeptide, and a pharmaceutically acceptable carrier. 1. A polypeptide for inhibiting , treating or diagnosing metastasis , which is a polypeptide , a derivative polypeptide , or a mutated polypeptide of a fibronectin-binding domain of dipeptidyl peptidase IV.2. The polypeptide as claimed in claim 1 , wherein the fibronectin-binding domain of dipeptidyl peptidase IV has a sequence with 70-100% similarity to a sequence represented by SEQ ID NO: 1.3. The polypeptide as claimed in claim 1 , wherein the fibronectin-binding domain of dipeptidyl peptidase IV has a sequence with 70-100% identity to a sequence represented by SEQ ID NO: 1.4. The polypeptide as claimed in claim 1 , wherein the fibronectin-binding domain of dipeptidyl peptidase IV has a sequence represented by SEQ ID NO: 1.5. The polypeptide as claimed in claim 1 , wherein the fibronectin-binding domain of dipeptidyl peptidase IV is a binding domain A of dipeptidyl peptidase IV.6. The polypeptide as claimed in claim 1 , wherein an N-terminal of the polypeptide is fused with a maltose-binding protein (MBP).7. An use of a polypeptide to prepare a pharmaceutical composition for inhibiting claim 1 , treating or diagnosing metastasis claim 1 , wherein the polypeptide is a polypeptide claim 1 , a derivative polypeptide claim 1 , or a mutated polypeptide of a fibronectin-binding domain of dipeptidyl peptidase IV.8. The use as claimed in claim 7 , wherein the fibronectin-binding domain of dipeptidyl peptidase IV has a sequence with 70-100% similarity to a sequence represented by SEQ ID NO: 1.9. The use as claimed in claim 7 , wherein the ...

MUTANT PROTEASES AND METHODS OF USE THEREOF

Mutant enzymes and methods of use thereof are provided. 1. A method for screening for modulators of an enzyme , said method comprisinga) contacting at least one mutant of said enzyme with at least one compound, wherein said mutant comprises at least one mutation in at least one blocking loop;b) measuring the activity of the mutant enzyme in the presence of said compound,wherein a modulation in the activity of the mutant enzyme in the presence of the compound compared to the activity of the mutant enzyme in the absence of the compound indicates that the compound is a modulator of the wild-type enzyme.2. The method of claim 1 , wherein said modulator is an inhibitor.3. The method of claim 1 , wherein said blocking loop is identified by protein structure.4. The method of claim 1 , wherein said mutation is in a tetra serine motif in said blocking loop.5. The method of claim 1 , wherein said enzyme is an isopeptidase.6. The method of claim 5 , wherein said isopeptidase is a deubiquitinating enzyme or ubiquitin-like protein (Ubl)-specific protease (Ulp).7. The method of claim 6 , wherein said enzyme is selected from the group consisting of ubiquitin specific protease 14 (USP14) claim 6 , USP24 claim 6 , USP42 claim 6 , USP36 claim 6 , USP53 claim 6 , USP26 claim 6 , USP10 claim 6 , USP51 claim 6 , SUMO1/sentrin specific protease 7 (SENP7) claim 6 , SENP1 claim 6 , and COP9 signalsome complex subunit 5 (CSN5).8. The method of claim 7 , wherein said enzyme is USP14.9. The method of claim 8 , wherein said mutant enzyme comprises an amino acid sequence having at least 80% homology with SEQ ID NO: 1 or 2 claim 8 , wherein at least one amino acid of the tetra serine motif is not a serine.10. The method of claim 9 , wherein said mutant enzyme comprises SEQ ID NO: 2.11. The method of claim 8 , wherein said wherein said mutant enzyme comprises an amino acid sequence having at least 80% homology with SEQ ID NO: 1 or 2 claim 8 , wherein at least one amino acid of the sequence ...

METHODS AND COMPOSITIONS FOR THE PRODUCTION OF EXTREMOPHILE ENZYMES FROM GREEN MICROALGAE AND CYANOBACTERIA

The present invention relates to compositions and methods for stable transformation of green microalgae and for production of transgenic green microalgae and/or cyanobacteria that produce extremophile enzymes as co-products during the growth of the green microalgae and/or cyanobacteria for lipid biofuel production. Thus, the present invention provides nucleic acid constructs and methods of transformation useful in the production of stably transformed green microalgae and/or cyanobacteria expressing extremophile enzymes in combination with lipid production for biofuel. 123-. (canceled)24. A method for producing one or more extremophile enzymes , the method comprising:(a) culturing a green microalgae cell, wherein the green microalgae cell is stably transformed with a heterologous nucleotide sequence encoding one or more extremophile enzymes and expresses the one or more extremophile enzymes; and(b) collecting the one or more extremophile enzymes from the green microalgae cell culture of (a), thereby producing one or more extremophile enzymes.25. The method of claim 24 , wherein the green microalgae cell is stably transformed with the heterologous nucleotide sequence encoding one or more extremophile enzymes claim 24 , bypropelling the heterologous nucleotide sequence at a green microalgae cell embedded in a gel at a velocity sufficient to pierce the cell wall, cell membrane and chloroplast membrane and deposit the heterologous nucleotide sequence within a chloroplast of the green microalgae cell;wherein the heterologous nucleotide sequence is incorporated into the chloroplast genome of the green microalgae cell, thereby producing a stably transformed green microalgae cell, andfurther wherein the heterologous nucleotide sequence is carried by a microprojectile and the heterologous nucleotide sequence is propelled at the green microalgae cell by propelling the microprojectile at the green microalgae cell.26. The method of claim 24 , wherein the heterologous nucleotide ...

METHOD FOR USING AN ENZYME RESISTANT TO HIGH PRESSURES

At least at least one embodiment of the present invention relates to a method for using a high pressure-resistant enzyme in a high pressure condition; a method for promoting the activity of the high pressure-resistant enzyme by means of a high pressure treatment; a composition, which contains the high pressure-resistant enzyme, for decomposing proteins under a high pressure condition; a composition, which contains the composition for decomposing proteins, for preparing natural flavoring substances; a container for high pressure treatment, which contains the composition for decomposing proteins; and a method for measuring the activity of the high pressure-resistant enzyme, which comprises a step of decomposing an azocasein solution serving as a substrate by using the high pressure-resistant enzyme treated under a high pressure condition. 1. A method for using an enzyme under a high pressure condition , wherein the enzyme is at least one selected from the group consisting of α-chymotrypsin , pepsin , trypsin , trypsin acetylated , flavourzyme , protease E and alcalase.2. A method for promoting the activity of at least one enzyme selected from the group consisting of α-chymotrypsin , pepsin , trypsin , trypsin acetylated , flavourzyme , protease E and alcalase , wherein the enzyme is treated at high pressure when heating.3. The method according to claim 1 , wherein the high pressure is 100 to 400 MPa.4. The method according to claim 1 , which is at least one selected from the group consisting of a method decomposing proteins claim 1 , carbohydrates or lipids claim 1 , a method for extracting bioactive compounds claim 1 , a method for modifying protein enzymes claim 1 , and a method for synthesizing functional ingredients with enzymes.5. The method according to claim 1 , wherein the high pressure is kept for 60 to 300 min.6. The method according to claim 1 , wherein the heating is conducted at 40° C. or higher for 2 min or longer.7. The method according to claim 2 , ...

ACE2-Fc FUSION PROTEINS FOR SARS-COV-2 MITIGATION

The present disclosure relates to recombinant fusion proteins comprising an extracellular domain of the human angiotensin-converting enzyme 2 (ACE2), optionally having altered amino acid residues that result in increased binding affinity for the S1 spike protein of SARS-CoV-2, linked to a human immunoglobulin Fc region, that can extend the protein half-life (T1/2) and/or the duration of action as a decoy receptor, and compositions and methods of use of these fusion proteins.

High-affinity human ace2 construct for use in diagnosing and treating coronaviruses

Provided herein, in some aspects, are polypeptide monomers comprising an angiotensin-converting enzyme 2 (ACE2) ectodomain and an oligomerization domain. Also provided herein are oligomeric complexes comprising ACE2 monomers. Methods of using such to monomers and oligomeric complexes for the diagnosis, prevention, and treatment of viral infections such as the coronavirus are also provided.

Collagen 7 and related methods

Disclosed are methods of making collagen 7, or functional fragments thereof, as well as collagen 7, and functional fragments thereof produced by such methods, nucleic acids encoding collagen 7, and functional fragments thereof, as well as vectors and host cells comprising such nucleic acids.

AMINOPEPTIDASE AND ITS USES

The present invention relates to the use of a TET protein as a N-terminus aromatic amino acid residues specific exopeptidase, said TET protein comprising the amino acid sequence as set forth in SEQ ID NO: 1. 1. A method for providing a N-terminus aromatic amino acid residues specific exopeptidase , wherein said a N-terminus aromatic amino acid residues specific exopeptidase is provided by a TET protein comprising , consisting essentially , or consisting of the amino acid sequence as set forth in SEQ ID NO: 1 ,or any homologous protein derived from said TET protein as set forth in SEQ ID NO: 1 by substitution, addition or deletion of at least one amino acid, provided that the derived protein retains at least 70% of identity with the amino acid sequence as set forth in SEQ ID NO: 1, and said derived protein retaining a N-terminus aromatic amino acid residues specific exopeptidase activity.2. A method for the modification of all or part of a polypeptide content of a substrate comprising peptides , polypeptides and/or proteins , wherein said modification is performed by at least a TET protein harboring at least a N-terminus aromatic amino acid residues specific exopeptidase activity , said at least a TET protein comprising , consisting essentially , or consisting of the amino acid sequence as set forth in SEQ ID NO: 1 ,or any homologous protein derived from said at least a TET protein as set forth in SEQ ID NO: 1 by substitution, addition or deletion of at least one amino acid, provided that the derived protein retains at least 70% of identity with the amino acid sequence as set forth in SEQ ID NO: 1, and said derived protein retaining a N-terminus aromatic amino acid residues specific exopeptidase activity.3. The method according to claim 1 , wherein said a TET protein or said derived protein originates from an extremophile microorganism belonging to the Methanococcales order.4Methanocaldococcus jannaschii.. The method according to claim 3 , wherein said extremophile ...

PSMA BINDING LIGAND-LINKER CONJUGATES AND METHODS FOR USING

Described herein are prostate specific membrane antigen (PSMA) binding conjugates that are useful for delivering therapeutic, diagnostic and imaging agents. Also described herein are pharmaceutical compositions containing them and methods of using the conjugates and compositions. Also described are processes for manufacture of the conjugates and the compositions containing them. 1. A conjugate comprising a ligand of PSMA (B) , a linker (L) , and a drug (D) , wherein the ligand includes one or more of a carbon-sulfur double bond , a phosphorus-sulfur double bond , a phosphorus-sulfur single bond , a thioester , or a combination thereof , and where the linker is covalently bound to the drug and the linker is covalently bound to the ligand , and where the linker comprises a chain of at least seven atoms.234-. (canceled)35. A kit comprising a sterile vial claim 1 , a composition comprising the conjugate of as a lyophilized solid claim 1 , and instructions describing use of the composition for treating a patient with an inflammatory disease claim 1 , wherein the vial is an amber glass vial with a rubber stopper and an aluminum tear-off seal claim 1 , and the vial is stored inside a cardboard box. This application is a continuation-in part of U.S. application Ser. No. 13/580,436, filed Aug. 22, 2012, which is a U.S. national application under 35 U.S.C. § 371(b) of International Application Serial No. PCT/US2011/026238 filed Feb. 25, 2011, and claims priority under 35 USC § 119(e) to U.S. Provisional Application Ser. No. 61/308,190, filed on Feb. 25, 2010, the entire disclosures of each of which are incorporated herein by reference.The invention described herein pertains to compounds and methods for treating diseases of the prostate, such as prostate cancer and related diseases. More specifically, embodiments of the invention described herein pertain to conjugates of biologically active agents conjugated to PSMA binding ligands.The prostate is one of the male reproductive ...

Anti-vegf protein compositions and methods for producing the same

The present disclosure pertains to compositions comprising anti-VEGF proteins and methods for producing such compositions.

Consensus Prostate Antigens, Nucleic Acid Molecule Encoding The Same And Vaccine And Uses Comprising The Same

Provided herein are consensus amino acid sequences of prostate antigens that are capable of breaking tolerance in a targeted species, including PSA, PSMA, STEAP and PSCA antigens. Also provided are nucleic acid sequences that encode one or more consensus amino acid sequences of prostate antigens PSA, PSMA, STEAP and PSCA, as well as genetic constructs/vectors and vaccines expressing the sequences. Also provided herein are methods for generating an autoimmune response against prostate cancer cells by administering one or more of the vaccines, proteins, and/or nucleic acid sequences that are provided.

COLLAGEN 7 AND RELATED METHODS

Disclosed are methods of making collagen 7, or functional fragments thereof, as well as collagen 7, and functional fragments thereof produced by such methods, nucleic acids encoding collagen 7, and functional fragments thereof, as well as vectors and host cells comprising such nucleic acids. 1. A method of making collagen 7 comprising:providing a cell recombinantly manipulated to express collagen 7, or a functional fragment thereof, and, optionally one or more polypeptides that increase expression of collagen 7, e.g., prolidase;culturing said cell under conditions sufficient for the production of collagen 7 and prolidase,thereby making collagen 7.2. The method of claim 1 , wherein said cell is genetically manipulated to express a glycosyl transferase.3. (canceled)4. The method of claim 1 , wherein said cell comprises exogenously introduced nucleic acid that encodes collagen 7 claim 1 , or a functional fragment thereof claim 1 , e.g. claim 1 , a high glycine codon optimized nucleic acid sequence described herein.5. The method of claim 1 , wherein said cell comprises exogenously introduced nucleic acid that encodes prolidase.6. The method of claim 1 , wherein said cell comprises exogenously introduced nucleic acid that encodes glycosyl transferase.7. The method of claim 1 , wherein said cell comprises an expression vector that comprises a sequence that encodes collagen 7.819-. (canceled)20. The method of claim 1 , further comprising recovering collagen 7 claim 1 , or functional fragment thereof claim 1 , from said cultured cell.2123-. (canceled)24. The method of claim 1 , wherein at least 30 claim 1 , 40 claim 1 , 50 claim 1 , 60 claim 1 , 70 claim 1 , 80 claim 1 , 90 or 95% of said collagen 7 claim 1 , or functional fragment thereof claim 1 , is incorporated into homotrimers.25. The method of claim 1 , wherein at least 30 claim 1 , 40 claim 1 , 50 claim 1 , 60 claim 1 , 70 claim 1 , 80 claim 1 , 90 or 95% of said collagen 7 claim 1 , or functional fragment thereof ...

Anti-vegf protein compositions and methods for producing the same

The present disclosure pertains compositions comprising anti-VEGF proteins and methods for producing such compositions.

ACTIVE LOW MOLECULAR WEIGHT VARIANTS OF ANGIOTENSIN CONVERTING ENZYME 2 (ACE2)

Disclosed are variants of ACE2, pharmaceutical compositions comprising the variants of ACE2, and treatment methods for reducing Angiotensin II (1-8) plasma levels and/or increasing Angiotensin (1-7) plasma levels in a subject in need thereof. The disclosed variants of ACE2 may include polypeptide fragments of ACE2 having ACE2 activity for converting AngII(1-8) to Ang(1-7). Suitable subjects suitable for the disclosed methods of treatment may include subjects having or at risk for developing diabetic and non-diabetic chronic kidney disease, acute renal failure and its prevention, chronic kidney disease, severe hypertension, scleroderma and its skin, pulmonary, kidney and hypertensive complications, malignant hypertension, renovascular hypertension secondary to renal artery stenosis, idiopathic pulmonary fibrosis, liver fibrosis such as in liver cirrhosis patients, an aortic aneurysm, cardiac fibrosis and remodeling, left ventricular hypertrophy, and an acute stroke. 119.-. (canceled)20. A method of delivering ACE2 activity to the kidney , the method comprising administering to a subject suffering from or at risk of developing kidney damage a pharmaceutical composition that delivers into the kidney an enzymatically active ACE2 variant that is sufficiently C-terminally truncated that it is susceptible to glomerular filtration.21. The method of claim 20 , wherein the subject is suffering from or at risk of kidney disease.22. The method of claim 21 , wherein the subject does not demonstrate symptoms of marked alterations in glomerular permeability.23. The method of claim 20 , wherein the subject is suffering from acute kidney injury.24. The method of claim 20 , wherein the subject is in early phase of diabetic kidney disease.25. The method of claim 20 , wherein the variant is a truncated human ACE2.26. The method of claim 20 , wherein the variant is truncated at a site C-terminal to its residue analogous to residue 522 of SEQ ID NO 2.27. The method of claim 26 , wherein ...

SOLUBILIZATION OF MSW WITH BLEND ENZYMES

The present invention relates to a method for solubilisation or hydrolysis of Municipal Solid Waste (MSW) with an enzyme blend and an enzyme composition for solubilization of Municipal Solid Waste (MSW), the enzyme composition comprising a cellulolytic background composition and a protease, lipase and/or beta-glucanase. 1. A process for solubilizing waste comprising:contacting Municipal Solid Waste (MSW) with an enzyme composition comprising a cellulolytic background composition comprising (a) a cellobiohydrolase I or variant thereof; (b) a cellobiohydrolase II or variant thereof; (c) a beta-glucosidase or variant thereof; and (d) a polypeptide having cellulolytic enhancing activity;and one or more enzymes selected from (i) a protease; (ii) a lipase and (iii) a beta-glucanase.2. (canceled)3. (canceled)4. The process of claim 1 , wherein the waste is pretreated.5. (canceled)6. (canceled)7. The process of claim 1 , wherein the enzyme composition further comprises one or more enzymes selected from (iv) a pectate lyase; (v) a mannanase; and (vi) an amylase.8. (canceled)9. (canceled)10Aspergillus fumigatusAspergillus fumigatusAspergillus fumigatusPenicillium. The process of claim 1 , wherein the cellobiohydrolase I of (a) is an cellobiohydrolase I or variant thereof; the cellobiohydrolase II of (b) is an cellobiohydrolase II or variant thereof; the beta-glucosidase of (c) is an beta-glucosidase or variant thereof; and the polypeptide having cellulolytic enhancing activity of (d) is a sp. GH61 polypeptide having cellulolytic enhancing activity; or homologs thereof.11Bacillus.. The process of claim 1 , wherein the protease (i) of the enzyme composition is derived from the genus12ThermomycesHumicola.. The process of claim 1 , wherein the lipase (ii) of the enzyme composition is derived from the genus or from the genus13Aspergillus.. The process of claim 1 , wherein the beta-glucanase (iii) of the enzyme composition is derived from a member of the genus14. The process of ...

METHODS FOR INCREASING GRAIN PRODUCTIVITY

The invention relates to methods for increasing plant yield, and in particular grain yield by reducing or abolishing the expression and/or activity of OTUB1 in a plant. Also described are genetically altered plants characterised by the above phenotype and methods of producing such plants. 1. A method of increasing grain yield in a plant , the method comprising reducing the expression of at least one nucleic acid encoding a otubain-like protease (OTUB1) and/or reducing the activity of OTUB1.2. (canceled)3. The method of claim 1 , wherein the method comprises introducing at least one mutation into at least one nucleic acid sequence encoding a OTUB1 polypeptide and/or the promoter of the OTUB1 polypeptide.4. (canceled)5. (canceled)6. The method of claim 1 , wherein the nucleic acid encodes a OTUB1 polypeptide wherein the OTUB1 polypeptide comprises SEQ ID NO: 1 or a functional homologue or variant thereof and wherein the nucleic acid encoding a OTUB1 promoter comprises SEQ ID NO: 6 or a functional variant or homologue thereof.7. (canceled)8. (canceled)9. (canceled)10. (canceled)11. The method of claim 1 , wherein the method comprises using RNA interference to reduce the expression of a least one OTUB1 nucleic acid.12. (canceled)13. (canceled)14. The method of claim 1 , wherein the plant is selected from rice claim 1 , wheat claim 1 , maize claim 1 , sorghum claim 1 , barley claim 1 , soybean claim 1 , and brassica.15. (canceled)16. A genetically altered plant claim 1 , part thereof or plant cell claim 1 , wherein said plant comprises at least one mutation in at least one nucleic acid encoding an OTUB1 polypeptide and/or the OTUB1 promoter or wherein the plant comprises an RNA interference construct that reduces the expression of an OTUB1 polypeptide.17. (canceled)18. (canceled)19. (canceled)20. (canceled)21. (canceled)22. (canceled)23. (canceled)24. (canceled)25. The genetically altered plant of claim 16 , wherein claim 16 , the nucleic acid encoding a OTUB1 ...

TREATMENT OF DISEASES RELATED TO HYPERACTIVITY OF THE COMPLEMENT SYSTEM

Raising the level of Factor I above physiological levels can be used to treat diseases in which the underlying pathology is linked to overactivity of the C3b-feedback cycle and the generation and pro-inflammatory effects of iC3b. Methods, agents, and compositions for treatment of such diseases are described. 173-. (canceled)74. A method of lowering the amount of C3b activity in a subject , the method comprising:increasing the level of C3b-inactivating activity and/or iC3b degrading activity in the subject;wherein the level of C3b-inactivating activity and/or iC3b degrading activity in the subject is increased to a level that exceeds a normal level of C3b-inactivating activity and/or iC3b degrading activity in the subject.75. The method according to claim 74 , further comprising not increasing the level of Factor H in the subject.76. The method according to claim 74 , wherein increasing the level of C3b-inactivating activity and/or iC3b degrading activity in the subject comprises increasing the level of Factor I in the subject.77. The method according to claim 76 , wherein the Factor I has at least 90% amino acid identity with native Factor I; andwherein the Factor I retains C3b-inactivating activity and/or iC3b degrading activity.78. The method according to claim 74 , wherein the level of C3b-inactivating activity and/or iC3b degrading activity is increased by at least 10% above the normal level.79. The method according to claim 74 , wherein the level of C3b-inactivating activity and/or iC3b degrading activity is increased by no more than 50% above the normal level.80. The method according to claim 74 , wherein the subject has a normal level of C3b-inactivating activity and/or iC3b degrading activity prior to increasing the level of C3b-inactivating activity and/or iC3b degrading activity in the subject.81. The method according to claim 74 , wherein the subject is human.82. The method according to claim 74 , wherein the subject suffers from Age-related Macular ...

Polypeptides Having Carboxypeptidase Activity And Polynucleotides Encoding Same

The present invention relates to isolated polypeptides having carboxypeptidase activity and isolated polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides. 1. A method of producing a hydrolysate from a proteinaceous substrate which comprises subjecting the substrate to an endopeptidase and a polypeptide having carboxypeptidase activity , selected from the group consisting of:(a) a polypeptide comprising an amino acid sequence having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 2;(b) a polypeptide encoded by a polynucleotide that hybridizes under at least very high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 1, (ii) the genomic DNA sequence comprising the mature polypeptide coding sequence of SEQ ID NO: 1, or (iii) the full-length complement of (i) or (ii), wherein very high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 μg/ml sheared and denatured salmon sperm DNA, and 50% formamide, and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 70° C.; and(c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1;wherein said polypeptide has the carboxypeptidase activity of SEQ ID NO: 2.2. The polypeptide of claim 1 , comprising the amino acid sequence of SEQ ID NO: 2 or the mature polypeptide thereof; or a fragment thereof having carboxypeptidase activity.3E. coli. The polypeptide of claim 1 , which is encoded by the polynucleotide contained in plasmid pTter44C2 which is contained in NRRL B-50207.4. A method of obtaining from a proteinaceous substrate a hydrolysate enriched in free glutamic acid and/or peptide bound glutamic acid residues claim 1 , comprising ...

Milling Process

Process for treating crop kernels, comprising the steps of: a) soaking kernels in water to produce soaked kernels; b) grinding the soaked kernels; c) treating the soaked kernels in the presence of an effective amount of a feruloyl esterase, wherein step c) is performed before, during or after step b). 2. The process of claim 1 , further comprising treating the soaked kernels in the presence of a protease.3. The process of claim 1 , further comprising treating the soaked kernels in the presence of an enzyme selected from the group consisting of an endoglucanase claim 1 , a xylanase claim 1 , a cellobiohydrolase I claim 1 , a cellobiohydrolase II claim 1 , a GH61 claim 1 , or a combination thereof.4. The process of claim 1 , further comprising treating the soaked kernels in the presence of an endoglucanase.5. The process of claim 1 , further comprising treating the soaked kernels in the presence of a xylanase.6. The process of claim 1 , wherein the kernels are soaked in water for about 2-10 hours.7. The process of claim 1 , wherein the soaking is carried out at a temperature between about 40° C. and about 60° C.8. The process of claim 1 , wherein the soaking is carried out at acidic pH.9. The process of claim 1 , wherein the soaking is performed in the presence of between 0.01-1%.10. The process of claim 1 , wherein the crop kernels are from corn (maize) claim 1 , rice claim 1 , barley claim 1 , sorghum bean claim 1 , or fruit hulls claim 1 , or wheat.11Aspergillus,Chaetomium,Humicola,Thielavia,Penicillium.. The process of claim 1 , wherein the feruloyl esterase is derived from a strain of the genus a strain of a strain of a strain of and/or a strain of12. (canceled) This application contains a Sequence Listing in computer readable form. The computer readable form is incorporated herein by reference.The present invention relates to an improved process of treating crop kernels to provide a starch product of high quality suitable for conversion of starch into mono- and ...

REWIRING ABERRANT CANCER SIGNALING TO A THERAPEUTIC EFFECTOR RESPONSE WITH A SYNTHETIC TWO-COMPONENT SYSTEM

Compositions and methods for targeted treatment of cancer are disclosed. In particular, the invention relates to methods of targeting anti-cancer therapy to cells exhibiting aberrant signaling associated with cancer pathogenesis by administering synthetic signaling proteins that couple detection of an oncogenic signal to release of therapeutic agents into cancerous cells. 1. A method for targeted treatment of a cancer associated with hyperactivity of a receptor tyrosine kinase , the method comprising:a) administering to a subject in need thereof a therapeutically effective amount of a first fusion protein comprising a protease connected to a phosphotyrosine binding (PTB) domain capable of binding to a phosphorylated tyrosine residue on the receptor tyrosine kinase; andb) administering a therapeutically effective amount of a second fusion protein comprising an SH2 domain connected to i) a substrate comprising a cleavage site recognized by the protease and ii) an anti-cancer therapeutic agent, wherein cleavage of the substrate at the cleavage site by the protease of the first fusion protein releases the anti-cancer therapeutic agent from the second fusion protein.2. The method of claim 1 , wherein the receptor tyrosine kinase is a hyperactive ErbB receptor tyrosine kinase.3. The method of claim 1 , wherein the protease is a hepatitis C virus (HCV) NS3 protease.4. The method of claim 1 , wherein the first fusion protein further comprises a degron claim 1 , wherein degradation activity of the degron is inhibited by binding of the PTB domain of the fusion protein to the phosphorylated tyrosine residue on the receptor tyrosine kinase such that the fusion protein accumulates preferentially in cancerous cells.5. The method of claim 4 , wherein the degron is located in a loop of the PTB domain.6. The method of claim 4 , wherein the degron is a HIF1a degron.7. The method of claim 1 , wherein the PTB is a Shc PTB.8. The method of claim 1 , wherein the SH2 domain is a Vav1 SH2 ...

Method for Producing an Acidified Milk Drink

The present invention relates to a method for producing an acidified milk drink using an enzyme which reduces the isoelectric point of the milk proteins. The invention also relates to a novel enzyme having deamidase activity and its use in production of an acidified milk drink.

ENHANCED PROTEIN EXPRESSION

The present invention relates in general to bacterial cells having a genetic alteration that results in increased expression of a protein of interest and methods of making and using such cells. Aspects of the present invention include Gram positive microorganisms, such as species, having a genetic alteration that reduces the expression of a gene in the pdh operon and results in enhanced expression of a protein of interest. 1Bacillus. A method for increasing expression of a protein of interest from a sp. cell comprising:{'i': 'Bacillus', 'a) obtaining an altered sp. cell capable of producing a protein of interest, wherein said altered cell comprises at least one genetic alteration that reduces expression of one or more genes in the pdh operon; and'}{'i': 'Bacillus', 'b) culturing said altered cell under conditions such that said protein of interest is expressed by said altered cell, wherein expression of said protein of interest is increased in said altered cell compared to the expression of said protein of interest in a corresponding unaltered sp. cell grown under essentially the same culture conditions.'}2. (canceled)3Bacillus. The method of claim 1 , wherein said altered cell has reduced expression of the pdhA gene as compared to the expression of the phdA gene in a corresponding unaltered sp. cell grown under essentially the same culture conditions.4Bacillus. The method of claim 1 , wherein said altered cell has reduced expression of the pdhB gene as compared to the expression of the phdB gene in a corresponding unaltered sp. cell grown under essentially the same culture conditions.5Bacillus. The method of claim 1 , wherein said altered cell has reduced expression of the pdhA gene and the pdhB gene as compared to the expression of the phdA gene and the pdhB gene in a corresponding unaltered sp. cell grown under essentially the same culture conditions.6. The method of claim 1 , wherein said genetic alteration results in a decrease in the level of an mRNA ...

DIGITAL MICROBIOLOGY

Methods, compositions, and kits are provided for rapidly analyzing microbial growth and/or number in a plurality of water-in-oil emulsion droplets. 190-. (canceled)91. A method for determining the presence or absence of a micro-organism in a sample , the method comprising:i) encapsulating a sample in a plurality of water-in-oil emulsion droplets wherein the water-in-oil emulsion droplets further encapsulate a microbiological growth medium;ii) incubating the plurality of water-in-oil emulsion droplets at a temperature permissive of microbiological growth, and for a period of time sufficient to allow the target microorganisms to go through 5 to 45 doubling times;iii) identifying water-in-oil emulsion droplets comprising target microorganisms; andiv) responsive to identifying the target microorganism in at least one water-in-oil emulsion droplet, determining that the target microorganism is present in the sample, wherein the target microorganisms are selected from yeasts and molds and wherein the incubating step is performed for a period of time corresponding to:at least about 6 hours and no more than about 12 hours when the target microorganisms are yeasts; andat least about 8 hours and no more than about 36 hours when the target microorganisms are molds.92. A method for determining the presence or absence of a micro-organism in a sample , the method comprising:i) encapsulating a sample in a plurality of water-in-oil emulsion droplets comprising an intercalating dye and no lysing agent wherein the water-in-oil emulsion droplets further encapsulate a microbiological growth medium;ii) identifying water-in-oil emulsion droplets comprising target microorganisms by detection of the intercalating dye; andiii) responsive to identifying the target microorganism in at least one water-in-oil emulsion droplet, determining that the target microorganism is present in the sample.93. The method of claim 92 , wherein:the water-in-oil droplet further comprises labeled protein;the ...

VECTOR LIBRARY FOR YEAST TWO HYBRID SCREENING AND METHOD FOR IDENTIFYING DEUBIQUITINATING ENZYME BINDING TO TARGET PROTEIN USING SAME

The present invention provides a vector library for yeast two-hybrid screening of a deubiquitinating enzyme that binds to a target protein and a method for identifying a deubiquitinating enzyme binding to a target protein using the same. Also, the present invention provides a method for screening an agent having anti-cancer activity targeting the deubiquitinating enzyme USP1, USP7, USP12, or USP49 identified by said identifying method. 1. A vector library for yeast two-hybrid screening of a deubiquitinating enzyme that binds to a target protein , comprising:a vector obtained by inserting a gene encoding a deubiquitinating enzyme USP1 in an empty vector having a DNA-binding domain;a vector obtained by inserting a gene encoding a deubiquitinating enzyme USP2 in an empty vector having a DNA-binding domain;a vector obtained by inserting a gene encoding a deubiquitinating enzyme USP3 in an empty vector having a DNA-binding domain;a vector obtained by inserting a gene encoding a deubiquitinating enzyme USP4 in an empty vector having a DNA-binding domain;a vector obtained by inserting a gene encoding a deubiquitinating enzyme USP5 in an empty vector having a DNA-binding domain;a vector obtained by inserting a gene encoding a deubiquitinating enzyme USP6 in an empty vector having a DNA-binding domain;a vector obtained by inserting a gene encoding a deubiquitinating enzyme USP7 in an empty vector having a DNA-binding domain;a vector obtained by inserting a gene encoding a deubiquitinating enzyme USP8 in an empty vector having a DNA-binding domain;a vector obtained by inserting a gene encoding a deubiquitinating enzyme USP10 in an empty vector having a DNA-binding domain;a vector obtained by inserting a gene encoding a deubiquitinating enzyme USP11 in an empty vector having a DNA-binding domain;a vector obtained by inserting a gene encoding a deubiquitinating enzyme USP12 in an empty vector having a DNA-binding domain;a vector obtained by inserting a gene encoding a ...

METHODS FOR CONTROLLING PROTEASE PRODUCTION

The present description is related to the field of protein production. It introduces novel host cells with low protease activity, a novel protease regulator, its use in expression systems and protein production, and a method of producing host cells for protein production 123-. (canceled)24. A host cell comprising at least one inactivated chromosomal gene wherein the inactivated chromosomal gene comprises a nucleic acid sequence encoding a polypeptide comprising a sequence having at least 90% sequence identity with the amino acids 402-533 of SEQ ID NO: 13;the inactivated chromosomal gene is inactivated by disruption, inhibition of translation or transcription of the chromosomal gene, at least partial deletion, truncation, deletion, insertion, mutation, or silencing, by RNAi, or by CRISPR/Cas9 technology;and the host cell has reduced protease activity compared to the host cell without said inactivation.25AscomycotaPezizomycotinaHypocrealesMicroascalesAspergillus, Chrysosporium, MyceliophthoraHumicolaTrichoderma, Hypocrea, Fusarium, Gibberella, Nectria, Stachybotrys, Claviceps, Metarhizium, Villosiclava, Ophiocordyceps, CephalosporiumScedosporiumTrichoderma reesei, Hypocrea jecorina, T. citrinoviridae, T. longibrachiatum, T. virens, T. harzianum, T. asperellum, T. atroviridae, T. parareesei, Fusarium oxysporum, F. gramineanum, F. pseudograminearum, F. venenatum, Gibberella fujikuroi, G. mondiformis, G. zeaea, Nectria haematococca, Stachybotrys chartarum, S. chlorohalonata, Claviceps purpurea, Metarhizium acridum, M. anisopliae, Villosiclava virens, Ophiocordyceps sinensis, Acremonium chrysogenum, Scedosporium apiospermum, Aspergillus niger, A. awamori, A. oryzae, Chrysosporium lucknowense, Myceliohpthora thermophila, Humicola insolensHumicola griseaTrichoderma reesei.. The host cell of claim 24 , wherein the host cell is selected from the group consisting of filamentous fungal cells from Division claim 24 , Subdivision ; preferably from the group consisting of members ...

PROSTATE-ASSOCIATED ANTIGENS AND VACCINE-BASED IMMUNOTHERAPY REGIMENS

The present disclosure provides (a) isolated immunogenic PAA polypeptides; (b) isolated nucleic acid molecules encoding immunogenic PAA polypeptides; (c) vaccine compositions comprising an immunogenic PAA polypeptide or an isolated nucleic acid molecule encoding an immunogenic PAA polypeptide; (d) methods relating to uses of the polypeptides, nucleic acid molecules, and compositions; and (e) vaccine-based immunotherapy regimens which involve co-administration of a vaccine in combination with an immune-suppressive-cell inhibitor and an immune-effector-cell enhancer. 126-. (canceled)27. A multi-antigen construct comprising two coding nucleotide sequences , wherein the two coding nucleotide sequences encode two different immunogenic PAA polypeptides selected from the group consisting of:(1) an immunogenic PSMA polypeptide and an immunogenic PSA polypeptide; and(2) an immunogenic PSA polypeptide and an immunogenic PSCA polypeptide.28. The multi-antigen construct according to claim 27 , wherein the immunogenic PSA polypeptide comprises an amino acid sequence selected from the group consisting of:(1) an amino acid sequence comprising amino acids 27-263 of SEQ ID NO: 15;(2) an amino acid sequence comprising amino acids 4-240 of SEQ ID NO:17; and(3) the amino acid sequence of SEQ ID NO:17.29. The multi-antigen construct according to claim 28 , wherein the immunogenic PSCA polypeptide comprises an amino acid sequence selected from the group consisting of:(1) the amino acid sequence of SEQ ID NO:21;(2) an amino acid sequence comprising amino acids 2-125 of SEQ ID NO:21; and(3) an amino acid sequence comprising amino acids 4-125 Of SEQ ID NO:21.30. The multi-antigen construct according to claim 28 , wherein the immunogenic PSMA polypeptide comprises an amino acid sequence selected from the group consisting of:(1) an amino acid sequence comprising amino acids 15-750 of SEQ ID NO:1;(2) the amino acid sequence of SEQ ID NO:3;(3) the amino acid sequence of SEQ ID NO:5;(4) the ...

ANGIOTENSIN-CONVERTING ENZYME 2 (ACE2) IMMUNOADHESIN MICROBODY

Provided are polypeptides comprising an enzymatically inactive angiotensin-converting enzyme 2 (ACE2) ectodomain, a segment of an immunoglobulin Fc and optionally a purification tag. A cDNA or an expression vector encoding the polypeptide along with a method of culturing cells comprising the expression vector is also provided. The disclosure also provides a method for prophylaxis or therapy for a Coronavirus infection by administering the polypeptide to an individual in need thereof. 1. A polypeptide comprising an enzymatically inactive angiotensin-converting enzyme 2 (ACE2) ectodomain that does not comprise an intact ACE2 transmembrane domain or an intact cytoplasmic tail , the polypeptide further comprising a segment of an immunoglobulin Fc that is not an intact Fc region.2. The polypeptide of claim 1 , wherein the ACE2 is enzymatically inactive by a mutation of an amino acid in an ACE2 catalytic active site.3. The polypeptide of claim 2 , wherein the mutation is at position 345 of SEQ ID NO:2 such that the amino acid at said position in not Histidine.4. The polypeptide of claim 3 , wherein the amino acid at position 345 of SEQ ID NO:2 is an alanine.5. The polypeptide claim 1 , wherein the segment of the immunoglobulin Fc that is not an intact Fc region comprises a microbody.6. The polypeptide of claim 5 , wherein the microbody comprises a segment of an Fc IgG-CH3.7. The polypeptide of claim 6 , wherein the microbody comprises SEQ ID NO:3.8. The polypeptide of claim 1 , wherein the polypeptide comprises the sequence of amino acids 1-871 of SEQ ID NO:5.9. The polypeptide of claim 8 , wherein the polypeptide further comprises a purification tag.10. The polypeptide of claim 9 , wherein the purification tag is at the C-terminus of the polypeptide.11. The polypeptide of claim 10 , wherein the purification tag comprises a poly-Histidine tag.12. A cDNA or an expression vector encoding the polypeptide of .13. A method comprising culturing cells comprising the expression ...

ANTIGENS OF PNEUMOCYSTIS MURINA AND USES THEREOF

A surface protein of the murine fungal pathogen can be used to generate an immune response in a recipient animal or human that provides prophylactic protection and an anti-fungal activity in subjects already infected with a species. Further, the disclosure provides novel polypeptides or peptides derived from the surface protein Surface Peptidase 1 (SPD-1) that are useful, alone or in combination with the SPD-1 polypeptide, in compositions and methods for the generation of an anti-immune reaction by a recipient subject. The compositions and methods of the disclosure provide advantageous alternatives to available immunogenic determinants for the treatment or prevention of fungal pneumonia. 1Pneumocystis murinaPneumocystis murinaPneumocystis. An immunogenic composition comprising at least one immunogenic component selected from the group consisting of: (a) an isolated Surface Peptidase 1 (SPD-1) polypeptide of and having an amino acid sequence having at least 90% similarity to the sequence according to SEQ ID NO: 1 and (b) at least one immunogenic fragment of an SPD-1 polypeptide of wherein said at least one fragment has an amino acid sequence of at least 90% similarity to an amino acid sequence selected from the group consisting of: SEQ ID NOs: 2 , 4 , 6 , 7 , and 10-40 , and wherein the composition further comprises a pharmaceutically acceptable carrier and is formulated to induce an anti-immune response when administered to an animal or human recipient.2. The composition of claim 1 , wherein the pharmaceutically acceptable carrier comprises an adjuvant.3. The composition of claim 1 , wherein the SPD-1 polypeptide has an amino acid sequence according to SEQ ID NO: 1.4. The composition of claim 1 , wherein the at least one immunogenic fragment of the SPD-1 polypeptide has an amino acid sequence selected from the group consisting of: SEQ ID NOs: 2 claim 1 , 4 claim 1 , 6 claim 1 , 7 claim 1 , and 10-40.5. The composition of claim 3 , wherein the at least one ...

METHODS OF AAV VECTOR PRODUCTION BY MODULATING DEUBIQUITINATING ENZYME ACTIVITY

Disclosed are host cells that have modulated DUB gene product activity and methods of using such host cells for increased AAV vector production yield. The disclosure also provides fusion proteins that comprise a DUB domain from a DUB gene product and a viral protein domain, and methods of increasing production of AAV vectors from a host cell that comprise expressing at least one of these fusion proteins in the host cell. 1. A host cell for producing AAV vectors comprising a modulated DUB gene product activity.2. The host cell of claim 1 , wherein the modulated DUB gene product activity comprises at least one of (a) alteration of expression of the DUB gene or (b) alteration of the subcellular localization of the DUB gene product.3. The host cell of claim 1 , wherein the modulated DUB gene product activity comprises expression of a DUB gene that is not normally expressed in the host cell.4. The host cell of claim 1 , wherein the modulated DUB gene product activity comprises overexpression of a DUB gene in the host cell.5. The host cell of claim 1 , wherein the modulated DUB gene product activity comprises suppression of expression of a DUB gene in the host cell.6. The host cell of claim 1 , wherein the modulated DUB gene product activity comprises a loss of the DUB gene activity in the host cell.7. The host cell of claim 1 , wherein the DUB gene is wild-type.8. The host cell of claim 1 , wherein the DUB gene is mutated.9. The host cell of claim 1 , wherein the modulated DUB gene product activity comprises alteration of the subcellular localization of the DUB gene product.10. The host cell of claim 9 , wherein the DUB gene product comprises a heterologous localization peptide.11. The host cell of claim 9 , wherein the DUB gene is wild-type and the DUB gene product is fused with a wild-type AAP protein.12. The host cell of claim 9 , wherein the DUB gene is mutant and the DUB gene product is fused with a wild-type AAP protein.13. The host cell of claim 9 , wherein the ...

Anti-Human Transferrin Receptor Antibody Permeating Blood-Brain Barrier

Disclosed are a means to convert compounds having physiological or pharmacological activity and unable to pass through the blood-brain barrier into a form that allows them to pass through the blood-brain barrier, and compounds converted thereby. The means is an anti-human transferrin receptor antibody and the converted compounds are molecular conjugates between physiologically active protein or pharmacologically active low-molecular-weight compounds and an anti-human transferrin receptor antibody. 169-. (canceled)70. A fusion protein comprising the amino acid sequences of a humanized anti-human transferrin receptor antibody and iduronate 2-sulfatase ,wherein the amino acid sequence of iduronate 2-sulfatase is linked to the light chain of the antibody on the C-terminal side or the N-terminal side of the light chain, or linked to the heavy chain of the antibody on the C-terminal side or the N-terminal side of the heavy chain, andwherein the light chain variable region and the heavy chain variable region of the antibody are selected from (1) or (2) below:(1) the light chain variable region comprising the amino acid sequence having an identity not lower than 90% to the amino acid sequence set forth as SEQ ID NO: 191, and comprising the amino acid sequence set forth as SEQ ID NO:16 as CDR1, the amino acid sequence set forth as SEQ ID NO:18 as CDR2, and the amino acid sequence set forth as SEQ ID NO:20 as CDR3, andthe heavy chain variable region comprising the amino acid sequence having an identity not lower than 90% to the amino acid sequence set forth as SEQ ID NO: 205, and comprising the amino acid sequence set forth as SEQ ID NO:88 as CDR1, the amino acid sequence set forth as SEQ ID NO:90 as CDR2, and the amino acid sequence set forth as SEQ ID NO:92 as CDR3;(2) the light chain variable region comprising the amino acid sequence having an identity not lower than 90% to the amino acid sequence set forth as SEQ ID NO: 191, and comprising the amino acid sequence set ...

Catalysis deactivated angiotensin-converting enzyme 2 (ACE2) variants and their uses

Angiotensin-converting enzyme 2 (ACE2) has been confirmed as a specific receptor for several (3 group coronaviruses include severe respiratory syndrome (SARS) coronavirus (SARS-CoV-1) and recently the causative agent for the World pandemic CoVID-19, SARS-CoV-2, and low pathogenic coronavirus of HCoV-NL63, a member in α-coronavirus group. Viral spike protein (S) of viral envelope is confirmed to bind to ACE2 as viral receptor to start a virus replication cycle. The present invention provides ACE2 and its mutants or variants, the viral or non-viral vectors thereof. Methods of treatment of viral infection of a human subject by using such mutants or variants are also provided.

Consensus Prostate Antigens, Nucleic Acid Molecule Encoding The Same And Vaccine And Uses Comprising The Same

Provided herein are consensus amino acid sequences of prostate antigens that are capable of breaking tolerance in a targeted species, including PSA, PSMA, STEAP and PSCA antigens. Also provided are nucleic acid sequences that encode one or more consensus amino acid sequences of prostate antigens PSA, PSMA, STEAP and PSCA, as well as genetic constructs/vectors and vaccines expressing the sequences. Also provided herein are methods for generating an autoimmune response against prostate cancer cells by administering one or more of the vaccines, proteins, and/or nucleic acid sequences that are provided. 1. A nucleic acid molecule comprising a coding sequence encoding one or more proteins selected from the group comprising:a) SEQ ID NO:2, a protein that is 98% homologous to SEQ ID NO:2, provided amino acids 69, 78, 80, 82, 102, 110, 137, 139, 165, 189, 203, 220, 232 and 248 of SEQ ID NO:2 are conserved, or an immunogenic fragment of SEQ ID NO:2 comprising amino acids corresponding to at least 256 amino acid residues of SEQ ID NO:2 provided amino acids 69, 78, 80, 82, 102, 110, 137, 139, 165, 189, 203, 220, 232 and 248 of SEQ ID NO:2 are conserved;b) SEQ ID NO:4, a protein that is 98% homologous to SEQ ID NO:4, provided amino acids 21, 86, 127, 129, 154, 156, 182, 195, 206, 218, 220, 237, 249, 255, 265, 271 or 275 of SEQ ID NO:4 are conserved, or an immunogenic fragment of SEQ ID NO:4 comprising amino acids corresponding to at least 274 amino acid residues of SEQ ID NO:4, provided amino acids 21, 86, 127, 129, 154, 156, 182, 195, 206, 218, 220, 237, 249, 255, 265, 271 or 275 of SEQ ID NO:4 are conserved;c) SEQ ID NO:6, a protein that is 98% homologous to SEQ ID NO:6, provided amino acids 14, 15, 32, 47, 58, 79, 111, 157, 223, 320, 350, 475, 499, 569, 613, 624, 653, 660, 663, 733 and 734 of SEQ ID NO:6 are conserved, or an immunogenic fragment of SEQ ID NO:6 comprising amino acids corresponding to at least 735 amino acid residues of SEQ ID NO:6, provided amino acids 14, 15 ...

Methods, Compositions, and Systems for Detecting Coronavirus Neutralizing Antibodies

The present disclosure relates to methods, compositions, and systems for detecting whether a subject exposed to a coronavirus has developed a neutralizing antibody response. Also disclosed are methods for determining whether a patient infected by a coronavirus is likely to respond to treatment with an antibody preparation. Also disclosed are methods for detecting the level of neutralizing antibody response in a sample of serum from a subject exposed to a coronavirus or to a coronavirus vaccine. 1. A method for detecting whether a subject exposed to a coronavirus has developed a neutralizing antibody response , comprising: i) a nucleic acid encoding a coronavirus spike protein, and', 'ii) a viral expression vector which comprises an indicator nucleic acid which produces a detectable signal;, '(a) transfecting into a plurality of first cells'}(b) incubating the first cells under conditions such that the first cells produce viral particles comprising the coronavirus spike protein;(c) contacting the viral particles of step (b) with a second cell in the presence or absence of a sample comprising an antibody from the subject, wherein the second cell expresses a cell surface receptor to which the coronavirus binds;(d) measuring the amount of the detectable signal produced by the second cell in the presence or absence of the sample; and(e) comparing the amount of signal measured in step (d) in the presence of the sample with the amount of signal produced in step (d) in the absence of the sample, wherein a reduced amount of signal measured in the presence of the sample indicates that the subject has developed a neutralizing antibody response capable of reducing infection.2. The method of claim 1 , wherein the indicator nucleic acid comprises an indicator gene.3. The method of claim 2 , wherein the indicator gene is a luciferase gene.4. The method of claim 1 , wherein the cell surface receptor is ACE-2.5. The method of claim 1 , wherein the subject was infected with severe ...

Selection methods for genetically-modified t cells

In some aspects, isolated transgenic cells (e.g., transgenic T cells) are provided that comprise or express a transgene and DHFRFS and/or TYMSSS. Methods for selecting transgeneic cells are also provided.

MOLECULES AND METHODS FOR ITERATIVE POLYPEPTIDE ANALYSIS AND PROCESSING

Reagents and methods for the digital analysis of proteins or peptides are provided. Specifically provided herein are proteins for identifying the N-terminal amino acid or N-terminal phosphorylated amino acid of a polypeptide. Also, an enzyme for use in the cleavage step of the Edman degradation reaction and a method for using this enzyme are described. 1. An isolated , synthetic , or recombinant N-terminal amino acid binding protein (NAAB) comprising:{'i': 'E. coli', '(1) an amino acid sequence having a glutamate residue at a position corresponding to position 42 of wild-type methionine aminopeptidase (eMAP) (SEQ ID NO: 1), a tryptophan residue at a position corresponding to position 46 of wild-type eMAP, a threonine or serine residue at a position corresponding to position 56 of wild-type eMAP, an aspartate residue at a position corresponding to position 57 of wild-type eMAP, a serine residue at a position corresponding to position 58 of wild-type eMAP, a leucine residue at a position corresponding to position 59 of wild-type eMAP, a threonine or serine residue at a position corresponding to position 60 of wild-type eMAP, a histidine residue at a position corresponding to position 62 of wild-type eMAP, an asparagine residue at a position corresponding to position 63 of wild-type eMAP, a isoleucine or valine residue at a position corresponding to position 65 of wild-type eMAP, an aspartate residue at a position corresponding to position 66 of wild-type eMAP, a glycine residue at a position corresponding to position 67 of wild-type eMAP, a histidine residue at a position corresponding to position 68 of wild-type eMAP, a glycine residue at a position corresponding to position 69 of wild-type eMAP, a serine or threonine residue at a position corresponding to position 70 of wild-type eMAP, a valine residue at a position corresponding to position 81 of wild-type eMAP, an arginine residue at a position corresponding to position 101 of wild-type eMAP, a histidine residue ...

Novel polypeptides with improved proteolytic stability, and methods of preparing and using same

The present invention includes methods of improving proteolytic stability of a polypeptide, comprising alkylating at least one selected from the group consisting of a N-terminus amino group, the NH group of the N-terminus first internal amide bond, another primary amino group, a thiol group and a thioether group within the polypeptide. The present invention further includes polypeptides incorporating such chemical modifications.

METHOD FOR CONTROLLING ENZYME PRODUCTIVITY OF MICROORGANISMS

An object of the present invention is to provide a novel method for controlling enzyme productivity of a microorganism. A pulsed electric field is applied to a microorganism to control the enzyme productivity of the microorganism. 1. A method for controlling the enzyme productivity of a microorganism characterized by applying a pulsed electric field to a microorganism.2. The method according to claim 1 , comprising a step of applying the pulsed electric field to the culture solution during culture of the microorganism.3. The method according to claim 2 , wherein the culture solution circulates in an electrode part that generates the pulsed electric field during culture.4. The method according to claim 2 , wherein the pulsed electric field is repeatedly applied during culture.5. The method according to claim 1 , wherein a pulse waveform of the pulsed electric field is a damped oscillation waveform.6. The method according to claim 1 , wherein a field strength of the pulsed electric field is 10 kV/cm to 50 kV/cm.7. The method according to claim 1 , wherein a production amount of one or more enzymes selected from the group consisting of amylase claim 1 , glucosidase claim 1 , galactosidase claim 1 , cellulase claim 1 , esterase claim 1 , lipase claim 1 , protease claim 1 , phosphatase claim 1 , peptidase claim 1 , nuclease claim 1 , deaminase claim 1 , oxidase claim 1 , dehydrogenase claim 1 , glutaminase claim 1 , pectinase claim 1 , catalase claim 1 , dextranase claim 1 , transglutaminase claim 1 , protein deamidase claim 1 , and pullulanase is controlled.8. The method according to claim 1 , wherein a production amount of one or more enzymes selected from the group consisting of α-amylase claim 1 , α-glucosidase claim 1 , β-glucosidase claim 1 , α-galactosidase claim 1 , β-galactosidase claim 1 , cellulase claim 1 , esterase claim 1 , lipase claim 1 , protease claim 1 , acid phosphatase claim 1 , alkaline phosphatase claim 1 , leucine peptidase claim 1 , alanine ...

FLAVOUR MODIFYING INGREDIENT DERIVED FROM DIETARY FIBRE

A method for making a flavour modifying ingredient, the method comprising subjecting a dietary fibre to enzymatic hydrolysis and/or fermentation; flavour modifying ingredients obtainable by said method; flavour compositions and food products comprises said flavour modifying ingredient; uses of said flavour modifying ingredient. 1Lactobacillus plantarum, Lactobacillus casei, Lactobacillus brevis, Lactobacillus helveticus, L. delbrueckiibulgaricus, Streptococcus thermophiles, Lactobacillus acidophilusBifidobacterium.. A method for making a flavour modifying ingredient , the method comprising subjecting a dietary fibre to enzymatic hydrolysis or fermentation , or enzymatic hydrolysis and fermentation , wherein the fermentation uses one or more than one lactic acid bacteria selected from the group consisting of ssp. and/or2. The method of claim 1 , wherein the dietary fibre is isolated dietary fibre.3. The method of claim 1 , wherein the dietary fibre is an aqueous slurry of dietary fibre.4. The method of claim 1 , wherein the dietary fibre is a cereal fibre claim 1 , a vegetable fibre claim 1 , or a fruit fibre.5. The method of claim 1 , wherein the enzymatic hydrolysis uses one or more enzymes selected from carbohydrases and proteolytic enzymes.6. The method of claim 1 , wherein the enzymatic hydrolysis uses at least one or more enzymes selected from cellulases claim 1 , pectinases claim 1 , and other carbohydrases.7. (canceled)8. The method of claim 1 , wherein the enzymatic hydrolysis is performed at a temperature ranging from about 25° C. to about 60° C.9. The method of claim 1 , wherein the enzymatic hydrolysis takes place for a period of time ranging from about 1 hour to about 48 hours.10. The method of claim 1 , wherein the fermentation is performed at a temperature ranging from about 20° C. to about 45° C.11. The method of claim 1 , wherein the fermentation takes place for a period of time ranging from about 1 day to about 10 days.12. (canceled)13. The method ...

Granule with hydrated barrier material

A granule having high stability and low dust is described. The granule includes a hydrated barrier material having moderate or high water activity. Also described are methods of producing the granules.

Cell Differentiation Marker and its Uses

The use of Dub3 protein, a nucleic acid molecule coding for the protein, or an inhibitor of the activity and/or of the expression of the protein for modulating cell differentiation. 1. A method for modulating cell differentiation comprising the administration to a determined cell:Dub3 protein, said protein comprising the amino acid sequence as set forth in SEQ ID NO: 1, or any variant thereof having at least 43% identity with said amino acid sequence SEQ ID NO: 1, and having ubiquitin hydrolase activity ora nucleic acid molecule coding for said protein or said variant thereof, oran inhibitor of the activity, i.e. the ubiquitin hydrolase activity and/or of the expression of said protein or said variant thereof.2. The method according to claim 1 , for modulating totipotent or pluripotent cell differentiation.3. A method for inducing dedifferentiation of differentiated cells claim 1 , the cells obtained from the dedifferentiation of differentiated cells being iPS cells claim 1 , the method comprising a step of administering to a differentiated cellsDub3 protein, said protein comprising the amino acid sequence as set forth in SEQ ID NO: 1, or any variant thereof having at least 43% identity with said amino acid sequence SEQ ID NO: 1, ora nucleic acid molecule coding for said protein or said variant thereof.4. The method according to for inducing dedifferentiation of differentiated cells claim 3 , wherein said cells Dub3 protein or said nucleic acid molecule coding for said protein are associated with at least an Oct family member protein and a Sox family member protein.5. The method according to claim 3 , wherein said Dub3 protein is expressed in said iPS cells at a level corresponding to at least 2 fold lower than the expression of said Dub3 protein in totipotent cell.6. A method for inducing a spontaneous differentiation of totipotent or pluripotent cells claim 3 , comprising the administration to a determined cell of an inhibitor of the activity and/or of the ...

PEPTIDASE AND ITS USES

The invention relates to the uses of a new characterized TET protein showed restricted to N-terminus glycine residues exopeptidase. The invention also relates to a method comprising said use of said new characterized TET protein as a N-terminus glycine residues specific exopeptidase. The invention further relates to a support wherein it is immobilized on said new characterized TET protein as a N-terminus glycine residues specific exopeptidase. 1. A method for providing a N-terminus glycine residues specific exopeptidase , wherein said N-terminus glycine residues specific exopeptidase is provided by a TET protein comprising the amino acid sequence as set forth in SEQ ID NO: 1 , or any homologous protein derived from said TET protein as set forth in SEQ ID NO: 1 by substitution , addition or deletion of at least one amino acid , provided that the derived protein retains at least 70% , preferably at least 79% of identity with the amino acid sequence as set forth in SEQ ID NO: 1 , and said derived protein retaining a N-terminus glycine residues specific exopeptidase activity.2. A method for the modification of all or part of the polypeptide content of a substrate comprising peptides , polypeptides and/or proteins harbouring a N-terminus glycine residue , wherein said modification is performed by at least a TET protein harbouring at least a N-terminus glycine residues specific exopeptidase activity , said at least TET protein comprising the amino acid sequence as set forth in SEQ ID NO: 1 ,or any homologous protein derived from said at least TET protein as set forth in SEQ ID NO: 1 by substitution, addition or deletion of at least one amino acid, provided that the derived protein retains at least 70%, preferably at least 79% of identity with the amino acid sequence as set forth in SEQ ID NO: 1, and said derived protein retaining a N-terminus glycine residues specific exopeptidase activity.3. The method according to claim 1 , wherein said TET protein or said derived ...

MOLECULES AND METHODS FOR ITERATIVE POLYPEPTIDE ANALYSIS AND PROCESSING

Reagents and methods for the digital analysis of proteins or peptides are provided. Specifically provided herein are proteins for identifying the N-terminal amino acid or N-terminal phosphorylated amino acid of a polypeptide. Also, an enzyme for use in the cleavage step of the Edman degradation reaction and a method for using this enzyme are described. 183-. (canceled)84. An isolated N-terminal amino acid binding protein (NAAB) , comprising a modified , non-naturally occurring tRNA synthetase (RS) that selectively binds to a N-terminal amino acid residue of a polypeptide with at least about a 1.5:1 ratio of specific to non-specific binding.85. The isolated NAAB of claim 84 , wherein the modified claim 84 , non-naturally occurring aminoacyl tRNA synthetase is coupled with or bound to a fluorescent label.86. The isolated NAAB of claim 85 , wherein the fluorescent label is covalently attached to the modified claim 85 , non-naturally occurring RS.87. The isolated NAAB of claim 84 , wherein the modified claim 84 , non-naturally occurring RS selectively binds to N-terminal amino acid residue of a particular type.88. The isolated NAAB of claim 87 , wherein the type of N-terminal amino acid residue is one selected from the group consisting of alanine claim 87 , arginine claim 87 , asparagine claim 87 , aspartic acid claim 87 , cysteine claim 87 , glutamine claim 87 , glutamic acid claim 87 , glycine claim 87 , histidine claim 87 , isoleucine claim 87 , leucine claim 87 , lysine claim 87 , methionine claim 87 , phenylalanine claim 87 , proline claim 87 , serine claim 87 , threonine claim 87 , tryptophan claim 87 , tyrosine claim 87 , and valine.89. The isolated NAAB of claim 84 , wherein the modified claim 84 , non-naturally occurring RS binds to an N-terminal amino acid residue with a post translational-modification.90. The isolated NAAB of claim 89 , wherein the N-terminal amino acid residue with a post translational-modification is a phosphorylated N-terminal amino acid ...

INFLAMMATION-ENABLING POLYPEPTIDES AND USES THEREOF

This present technology relates to the use of inflammation-enabling polypeptides (or their coding sequences) to screen for agents useful for the prevention, treatment and/or alleviations of symptoms associated with an inflammatory disorder, to identify individuals susceptible of developing an exacerbated inflammatory response as well as to determine if a therapeutic regimen is capable of preventing, treating or alleviating the symptoms associated to an inflammatory disorder in an individual. The present technology also provides methods for preventing, treating and/or alleviating the symptoms associated to an inflammatory condition based on the inhibition of expression or activity of the inflammation-enabling targets. 1. A method for assessing the ability of an agent to prevent , treat and/or alleviate the symptoms associated with an inflammatory condition in an individual , said method comprising:(a) combining the agent with an inflammatory enabling polypeptide selected from the group consisting of a USP15 polypeptide and a TRIM25 polypeptide;(b) measuring a biological activity of the inflammatory enabling polypeptide of step (a) to obtain a test level;(c) comparing the test level to a control level, wherein the control level is associated with the biological activity of the inflammatory enabling polypeptide observed during the onset or maintenance of the inflammatory condition; and(d) characterizing the agent as (i) useful for the prevention, treatment and/or alleviation of the symptoms associated with the inflammatory condition when the at least one biological activity associated with the test level is lower than the biological activity associated with the control level or (ii) lacking utility for the prevention, treatment and/or alleviation of the symptoms associated with the inflammatory condition when the at least one biological activity associated with the test level is equal to or higher than the biological activity associated with the control level.2. The ...

PROTEIN M FUSION PROTEINS AND USES

Fusion proteins with immunoglobulin binding properties, their uses and related methods and compositions are disclosed. The fusion proteins are comprised of an antibody-binding fragment of protein M from spp. conjugated to a receptor fragment. The receptor fragment is a protein fragment to which a pathogen or a toxin can specifically bind. The fusion proteins can be used to neutralize or eradicate a wide group of pathogens or toxins. 1. A method for neutralizing a pathogen , wherein the pathogen has a specific binding affinity for a receptor fragment , the method comprising: providing conditions for interaction between the pathogen and a fusion protein that comprises a polypeptide having at least 90% identity over its entire length with either the sequence set forth in SEQ ID NO: 1 or the sequence set forth in SEQ ID NO: 2 conjugated to the receptor fragment , whereby the fusion protein binds to and neutralizes the pathogen.2. The method according to claim 1 , wherein the receptor fragment is a protein fragment of a cellular receptor.3. The method according to claim 2 , wherein the pathogen is SARS-CoV-2 virus and the receptor fragment comprises the sequence set forth in SEQ ID NO: 15.4. The method according to claim 1 , wherein the fusion protein further comprises a spacer between the polypeptide and the receptor fragment.5. The method according to claim 1 , wherein the receptor fragment comprises one of the following sequences: SEQ ID NO: 16-36.6. The method according to claim 1 , wherein the fusion protein neutralizes the pathogen via recruitment of C1q protein.7. A method for eradicating a bloodborne pathogen in a subject claim 1 , wherein the pathogen has a specific binding affinity for a receptor fragment inside a body of the subject claim 1 , the method comprising:receiving a sample of blood, serum or plasma from the subject or from a donor compatible with the subject, wherein the sample comprises immunoglobulins;adding a fusion protein that comprises a ...

COMPOSITIONS AND METHODS COMPRISING KLK3 OR FOLH1 ANTIGEN

The present invention provides KLK3 peptides, FOLH1 peptides, recombinant polypeptides comprising same, recombinant nucleotide molecules encoding same, recombinant strains comprising same, and immunogenic and therapeutic methods utilizing same. 150-. (canceled)51. A method of inhibiting prostate cancer metastasis in a subject , comprising:{'i': 'Listeria', 'administering to the subject a composition comprising a recombinant strain encoding a recombinant fusion peptide consisting of a KLK3 peptide operatively linked to a non-KLK3 peptide, wherein the non-KLK3 peptide is a N-terminal non-hemolytic listeriolysin (LLO) peptide, the fusion peptide consisting of the sequence of SEQ ID NO: 54 or a sequence at least 99% homologous (throughout the length of the peptide), wherein the metastasis is inhibited.'}52ListeriaListeria.. The method of claim 51 , wherein the recombinant strain is an auxotrophic53Listeria. The method of claim 52 , wherein the auxotrophic strain is a dal/dat mutant comprising a deletion in the endogenous ActA gene.54ListeriaListeria. The method of claim 52 , wherein the auxotrophic strain comprises an episomal expression vector encoding a metabolic enzyme that complements the auxotrophy of the auxotrophic strain.55. The method of claim 54 , wherein the metabolic enzyme is an alanine racemase enzyme.56. The method of claim 54 , wherein the metabolic enzyme is a D-amino acid transferase enzyme.57ListeriaListeria monocytogenes. The method of claim 51 , wherein the recombinant is a recombinant strain.58. The method of claim 51 , wherein the inhibiting prostate cancer metastasis comprises metastasis-free prostate cancer progression.59. The method of claim 51 , wherein the inhibiting prostate cancer metastasis comprises a reduced rate of growth or reduced number of prostate cancer metastases.60. A method of treating prostate cancer metastasis in a subject claim 51 , comprising:{'i': 'Listeria', 'administering to the subject a composition comprising a ...

ENGINEERED PROTEASE VARIANTS

The present invention provides engineered protease polypeptides and compositions thereof. The engineered protease polypeptides have been optimized to provide improved activity, improved thermostability, protease stability, autolytic stability, and stability under a range of pH conditions, including acidic (pH<7) and basic (pH>7) conditions. The invention also relates to the use of the compositions comprising the engineered protease polypeptides for therapeutic and/or nutritional purposes. The present invention also provides polynucleotides encoding the engineered protease polypeptides, as well as methods for making the engineered polynucleotides and protease polypeptides. 1. A recombinant protease comprising an amino acid sequence comprising at least 70% , at least 75% , at least 80% , at least 85% , at least 90% , at least 91% , at least 92% , at least 93% , at least 94% , at least 95% , at least 96% , at least 97% , at least 98% , or at least 99% sequence identity to SEQ ID NO: 2 , 4 , 190 , 292 , 342 , 382 , 396 , 400 , 454 , 562 , 638 , 680 , 756 , 812 , 1030 , 1136 , 1180 , 1250 , 1308 , 1366 , 1370 , 1424 , 1532 , 1608 , 1650 , 1726 , 1782 , 2000 , 2106 , 2150 , 2220 , and/or 2278.2. (canceled)3. The recombinant protease of claim 1 , wherein said recombinant protease comprises a polypeptide sequence having at least 85% claim 1 , 86% claim 1 , 87% claim 1 , 88% claim 1 , 89% claim 1 , 90% claim 1 , 91% claim 1 , 92% claim 1 , 93% claim 1 , 94% claim 1 , 95% claim 1 , 96% claim 1 , 97% claim 1 , 98% claim 1 , 99% claim 1 , or more sequence identity to SEQ ID NO: 2 claim 1 , wherein said recombinant protease comprises at least one substitution at one or more positions selected from 236 claim 1 , 258 claim 1 , 261 claim 1 , 339 claim 1 , 439 claim 1 , 446 claim 1 , and 454 claim 1 , wherein the amino acid positions of said polypeptide sequence are numbered with reference to SEQ ID NO: 2.4. The recombinant protease of claim 3 , wherein the substitution at positions ...

COMPOSITION FOR CONVERTING INSULIN PRECURSOR INTO INSULIN ENZYME AND METHOD FOR CONVERTING INSULIN PRECURSOR INTO INSULIN BY USING SAME

The present invention relates to: a composition for an enzymatic conversion reaction for converting proinsulin or a proinsulin analogue into insulin or an insulin analogue, the composition comprises one or more selected from the group consisting of calcium chloride, glycine, and proline, and trypsin and/or carboxypeptidase B; and a method for converting proinsulin or a proinsulin analogue into insulin or an insulin analogue by using the composition. The present invention increases the structural stability of an insulin protein and the stability of the enzyme at the same time so as to significantly improve the enzymatic conversion yield and the purity of insulin, thereby being useful in insulin production processes. 1. A composition for enzymatically converting an insulin precursor into insulin , comprising:one or more selected from the group consisting of calcium chloride, glycine, and proline; andtrypsin and/or carboxypeptidase B.2. The composition according to claim 1 , further comprising a buffer solution.3. The composition according to claim 2 , wherein the buffer solution is 1-100 mM Tris-HCl or 1-100 mM borate.4. The composition according to claim 1 , wherein the trypsin is comprised at a weight ratio of 1/1 claim 1 ,000 to 1/40 claim 1 ,000 relative to the insulin precursor claim 1 , and/or the carboxypeptidase B is comprised at a weight ratio of 1/600 to 1/20 claim 1 ,000 relative to the insulin precursor.5. The composition according to claim 1 , wherein the insulin is insulin glargine.6. The composition according to claim 1 , wherein the composition comprises calcium chloride claim 1 , glycine claim 1 , and proline.7. The composition according to claim 6 , wherein calcium chloride is comprised at 5-30 mM claim 6 , glycine is comprised at 3-60 mM claim 6 , and proline is comprised at 20-80 mM.8. A method for converting an insulin precursor into insulin claim 1 , comprising adding an insulin precursor to the composition according to and carrying out a ...

Polynucleotide Constructs For In Vitro and In Vivo Expression

The present invention relates to polynucleotide constructs for in vitro and in vivo transcription/translation of genes of interest or variants of a gene of interest as well as microorganism host cells comprising such constructs and methods for producing a polypeptide of interest in such microorganism host cells.

DPEP-1 Binding Compositions and Methods of Use

Pharmaceutical compositions and methods of their use are provided for reducing inflammation in a subject, blocking leukocyte recruitment, inhibiting tumor metastasis, treating sepsis and preventing/reducing acute kidney injury. 1. A method for treating inflammation in a subject in need thereof , comprising administering an effective amount of a compound that binds to DPEP-1 to the subject , thereby treating inflammation.2. The method of claim 1 , wherein the compound is selected from a competitive antagonist claim 1 , a non-competitive antagonist claim 1 , and uncompetitive antagonist claim 1 , or a silent antagonist of DPEP-1 or a combination thereof.3. The method of claim 1 , wherein the compound comprises a peptide.4. The method of claim 3 , wherein the peptide comprises an LSALT sequence (LSALTPSPSWLKYKAL).5. The method of claim 3 , wherein the peptide comprises a GFE tripeptide sequence selected from CGFECVRQCPERC (GFE-1) or CGFELETC (GFE-2).6. The method of claim 5 , wherein the peptide is not conjugated to cilastatin.7. The method of claim 1 , wherein the compound comprises a blocking antibody of DPEP-1.8. The method of claim 1 , wherein the compound comprises a small molecule.10. The method of claim 1 , wherein the compound is provided as a pharmaceutical composition.11. The method of claim 10 , wherein the pharmaceutical composition is suitable for parenteral or intravenous administration.12. The method of claim 1 , wherein the effective amount is between about 0.01 mg/kg and about 100 mg/kg.13. The method of claim 1 , wherein the inflammation is associated with acute kidney injury.14. The method of claim 1 , further comprising identifying the subject by performing a diagnostic test to determine a need for reduction in inflammation.15. A method for block leukocyte recruitment in a subject in need thereof claim 1 , comprising administering an effective amount of a composition that binds to DPEP-1 to the subject claim 1 , thereby blocking leukocyte ...

Erap1-derived peptide and use thereof

Provided is a novel cancer-treating agent which can be used as a novel choice for the treatment of cancer. Specifically provided are: a peptide that inhibits binding of ERAP1 polypeptide to PHB2 polypeptide, which comprises a binding site of the ERAP1 polypeptide to the PHB2 polypeptide, and a pharmaceutical composition comprising the peptide. In addition, provided is a method for screening a drug candidate for treating and/or preventing cancer using inhibition of the binding of the ERAP1 polypeptide to PP1α polypeptide, PKA polypeptide or PKB polypeptide as an index.

Anti-vegf protein compositions and methods for producing the same

The present disclosure pertains to compositions comprising anti-VEGF proteins and methods for producing such compositions.

PSMA BINDING LIGAND-LINKER CONJUGATES AND METHODS FOR USING

Described herein are prostate specific membrane antigen (PSMA) binding conjugates that are useful for delivering therapeutic, diagnostic and imaging agents. Also described herein are pharmaceutical compositions containing them and methods of using the conjugates and compositions. Also described are processes for manufacture of the conjugates and the compositions containing them. 1. (canceled)235-. (canceled)38. The conjugate of claim 36 , wherein the ligand is a prostate specific membrane antigen (PSMA) that is a thiourea of two amino acids claim 36 , wherein the two amino acids are independently selected from asparagine claim 36 , aspartic acid claim 36 , cysteine claim 36 , glutamic acid claim 36 , lysine claim 36 , glutamine claim 36 , arginine claim 36 , serine claim 36 , ornithine claim 36 , threonine claim 36 , and combinations thereof.39. The conjugate of claim 36 , or a salt thereof claim 36 , wherein the chelating group is chelated to a radioactive metal isotope.40. The conjugate of claim 36 , or a salt thereof claim 36 , wherein the divalent linker is covalently bound to the chelating group through the formation of an amide bond claim 36 , and the divalent linker is covalently bound to the ligand through the formation of an amide bond.41. The conjugate of claim 36 , or a salt thereof claim 36 , wherein L is covalently bound to B through an amide bond.42. The conjugate of claim 36 , or a salt thereof claim 36 , wherein L is covalently bound to the chelating group through a carbon-nitrogen single bond.43. The conjugate of claim 37 , or a salt thereof claim 37 , wherein the chelating group is chelated to a radioactive metal isotope.44. The conjugate of claim 37 , or a salt thereof claim 37 , wherein the divalent linker is covalently bound to the chelating group through the formation of an amide bond claim 37 , and the divalent linker is covalently bound to the ligand through the formation of an amide bond.45. The conjugate of claim 37 , or a salt thereof ...

Immunogenic Fusion Proteins For The Treatment Of Cancer

Provided herein are compositions and methods for eliciting an immune response in a subject. In particular, the present disclosure is directed to immunogenic fusion proteins and methods of eliciting an immune response using host cells comprising nucleic acid molecules encoding said fusion proteins. 1195.-. (canceled)196. A recombinant nucleic acid molecule encoding three separate fusion proteins , wherein the three separate fusion proteins are EGFRvIIIx5-SSX2 , EGFRvIIIx5-PAP , and EGFRvIIIx5-NKX3.1(R41G)-PSMA , and wherein:a) EGFRvIIIx5-SSX2 is encoded by a nucleic acid sequence comprising the sequence set forth in SEQ ID NO:3;{'sub': '33-386', 'b) EGFRvIIIx5-PAPis encoded by a nucleic acid sequence comprising the sequence set forth in SEQ ID NO:11; and'}{'sub': 11-234', '1-20, 44-138, 169-750, 'c) EGFRvIIIx5-NKX3.1(R41G)-PSMAis encoded by a nucleic acid sequence comprising the sequence set forth in SEQ ID NO:15.'}197. The nucleic acid molecule of claim 196 , further comprising a promoter claim 196 , a signal sequence claim 196 , or both claim 196 , wherein the promoter claim 196 , signal sequence claim 196 , or both are operably linked with the nucleotide sequence encoding the EGFRvIIIx5 polypeptide and the nucleotide sequence encoding the SSX2 polypeptide.198. The nucleic acid molecule of which is operably linked to an ActA promoter claim 196 , said ActA promoter having the sequence set forth in SEQ ID NO:21.199. A method of eliciting an immune response in a subject claim 196 , said method comprising administering to the subject a therapeutically effective amount of a composition comprising a recombinant nucleic acid molecule claim 196 , wherein the nucleic acid molecule encodes the fusion proteins EGFRvIIIx5-SSX2 claim 196 , EGFRvIIIx5-PAP claim 196 , and EGFRvIIIx5-NKX3.1(R41G)-PSMA claim 196 , and wherein:a) EGFRvIIIx5-SSX2 is encoded by a nucleic acid sequence comprising the sequence set forth in SEQ ID NO:3;{'sub': '33-386', 'b) EGFRvIIIx5-PAPis encoded by a ...

Methods of identifying genetic variants

The present invention relates to identification of an abnormal splice site. Provided are methods of identifying an abnormal splice site. Methods of classifying the risk of abnormal splicing of a splice site are also provided. Databases for use in the methods provided herein are also disclosed.

NOVEL METALLOPROTEASES

Aspects of the present compositions and methods relate to novel metalloproteases, polynucleotides encoding the novel metalloproteases, and compositions and methods for use thereof. 1. A polypeptide comprising an amino acid sequence having at least 60% sequence identity to the amino acid sequence selected from the group consisting of SEQ ID NOs: 3 , 8 , 13 , 18 , 23 , 28 , 33 and 38.2. The polypeptide of claim 1 , wherein said polypeptide has at least 80% sequence identity to the amino acid sequence selected from the group consisting of SEQ ID NOs: 3 claim 1 , 8 claim 1 , 13 claim 1 , 18 claim 1 , 23 claim 1 , 28 claim 1 , 33 and 38.3. The polypeptide of any of or claim 1 , wherein said polypeptide has at least 95% sequence identity to the amino acid sequence selected from the group consisting of SEQ ID NOs: 3 claim 1 , 8 claim 1 , 13 claim 1 , 18 claim 1 , 23 claim 1 , 28 claim 1 , 33 and 38.4. The polypeptide of any of the above claims claim 1 , wherein said amino acid sequence is the amino acid sequence selected from the group consisting of SEQ ID NOs: 3 claim 1 , 8 claim 1 , 13 claim 1 , 18 claim 1 , 23 claim 1 , 28 claim 1 , 33 and 38.5. The polypeptide of any of the above claims claim 1 , wherein said polypeptide is derived from a member of the order Bacillales.6. The polypeptide of any of the above claims claim 1 , wherein said Bacillales member is a Paenibacillaceae family member.7Paenibacillus. The polypeptide of claim 6 , wherein said Bacillales member is a spp.8Planococcus. The polypeptide of any of - claim 6 , wherein said polypeptide is derived from a species.9. The polypeptide of any of the above claims claim 6 , wherein said polypeptide has protease activity.10. The polypeptide of claim 9 , wherein said protease activity comprises casein hydrolysis claim 9 , collagen hydrolysis claim 9 , elastin hydrolysis claim 9 , keratin hydrolysis claim 9 , soy protein hydrolysis or corn meal protein hydrolysis.11. The polypeptide of any of the above claims claim 9 ...

Multivalent particles compositions and methods of use

Provided herein are multivalent particles and compositions of multivalent particles for blocking viral infection.

Multiple Proteases Deficient Filamentous Fungal Cells and Methods of Use Thereof

The present disclosure relates to compositions and methods useful for the production of heterologous proteins in filamentous fungal cells. 116-. (canceled)17Myceliophthora thermophilaMyceliophthora thermophila. A filamentous fungal cell comprising at least one endogenous amp or pep protease having no protease activity and a recombinant polynucleotide encoding a mammalian polypeptide , wherein a gene encoding an endogenous amp protease having at least 98% identity to SEQ ID NO: 767 , 774 or 780 or pep protease having at least 98% identity to XP_003667167.1 of the filamentous fungal cell comprises a mutation that eliminates the activity of the endogenous amp or pep protease.18Myceliophthora thermophila. The filamentous fungal cell of claim 17 , wherein the gene encoding endogenous amp or pep protease is deleted.19Myceliophthora thermophila. The cell of comprising at least three claim 17 , four claim 17 , five claim 17 , six claim 17 , seven or eight endogenous proteases having eliminated activity.20Myceliophthora thermophila. The cell of claim 19 , wherein the proteases are selected from the group consisting of aspartic proteases claim 19 , trypsin-like serine proteases claim 19 , subtilisin proteases claim 19 , glutamic proteases claim 19 , and metalloproteases.21Myceliophthora thermophilaMyceliophthora thermophila. The cell of claim 19 , wherein the cell has no detectable protease activity of pep4 protease having at least 98% identity to SEQ ID NO: 499 claim 19 , slp2 protease having at least 98% identity to SEQ ID NO: 540 claim 19 , and slp3 protease having at least 98% identity to SEQ ID NO: 546.22Myceliophthora thermophila. The cell of claim 17 , wherein the mammalian polypeptide is selected from the group consisting of an antibody claim 17 , a growth factor claim 17 , an interferon claim 17 , a cytokine claim 17 , and an interleukin.23Myceliophthora thermophilaMyceliophthora thermophila. The cell of claim 17 , wherein the cell further comprises ALG3 having ...

DIPEPTIDYL PEPTIDASE-IV (DPPIV), INHIBITORY PEPTIDE COMPOUND, COMPOSITION CONTAINING THE SAME, AND PRODUCTION METHOD FOR THE SAME

A peptide compound having a dipeptidyl peptidase-IV (DPPIV) inhibitory activity or a composition containing the peptide compound that can make a contribution to the prevention of the onset of pathology or the progression in diabetes mellitus patients or those at risk of diabetes mellitus can be provided according to the present invention by a simple method using, as a raw material, milt of a fishery product, which has been eaten for ages and has high safety. In the present invention, a peptide compound having a peptidyl peptidase-IV (DPPIV) inhibitory activity obtained in a hydrolysate of a milt protein source obtained from a fishery product is used as an active component of a composition for inhibiting DPPIV. 111-. (canceled)12. A production method for a hydrolysate having a dipeptidyl peptidase-IV (DPPIV) inhibitory activity , comprising:hydrolyzing a protein source derived from milt of a fishery product to obtain the hydrolysate having a DPPIV inhibitory activity.1317.-. (canceled)18. The production method according to claim 12 , wherein the hydrolyzing step is performed under a condition such that at least one peptide compound selected from the group consisting of Phe-Pro-Val-Gly and salts thereof claim 12 , Ile-Pro-Leu and salts thereof claim 12 , Leu-Pro-Val-Leu and salts thereof claim 12 , and Val-Pro-Phe-Pro and salts thereof is contained in the hydrolysate.19. (canceled)20. A production method for a peptide compound having a dipeptidyl peptidase-IV (DPPIV) inhibitory activity claim 12 , comprising the steps of:{'claim-ref': {'@idref': 'CLM-00012', 'claim 12'}, '(a) obtaining a hydrolysate by the production method according to ; and'}(b) isolating the peptide compound having a DPPIV inhibitory activity from the hydrolysate.21. The method of claim 12 , wherein the fishery product is one or more of salmon claim 12 , pink salmon claim 12 , herring claim 12 , pacific cod claim 12 , skipjack claim 12 , yellowtail (young yellowtail) and squid.22. The method of ...

COMPOSITIONS AND METHODS COMPRISING KLK3 OR FOLH1 ANTIGEN

The present invention provides KLK3 peptides, FOLH1 peptides, recombinant polypeptides comprising same, recombinant nucleotide molecules encoding same, recombinant strains comprising same, and immunogenic and therapeutic methods utilizing same. 1Listeria. A recombinant strain expressing a folate hydrolase 1 (FOLH1) peptide , wherein either (a) the sequence of said FOLH1 peptide is a sequence selected from the group consisting of SEQ ID No: 41 , 43 , 44 , and 45; or (b) said FOLH1 peptide is an immunogenic fragment of a larger FOLH1 peptide , wherein the sequence of said larger FOLH1 peptide is selected from the group consisting of SEQ ID No: 41 , 43 , 44 , and 45.2Listeria. The recombinant strain of claim 1 , wherein said FOLH1 peptide is in the form of a fusion peptide claim 1 , wherein said fusion peptide further comprises a non-FOLH1 peptide claim 1 , wherein said non-FOLH1 peptide enhances the immunogenicity of said fragment.3Listeria. The recombinant strain of claim 1 , wherein said non-FOLH1 peptide is selected from the group consisting of a listeriolysin (LLO) peptide claim 1 , an ActA peptide claim 1 , and a PEST-like sequence peptide.4Listeria. The recombinant strain of claim 1 , wherein said FOLH1 peptide does not contain an FOLH1 signal sequence.5Listeria. An immunogenic composition comprising the recombinant strain of and an adjuvant.6ListeriaListeriaListeria monocytogenes. The recombinant strain of claim 1 , wherein said recombinant strain is a recombinant strain.7ListeriaListeria. The recombinant strain of claim 1 , wherein said recombinant strain has been passaged through an animal host.8ListeriaListeriaListeria. The recombinant strain of claim 1 , wherein said strain is an auxotrophic strain.9ListeriaListeria. The recombinant strain of claim 8 , wherein said auxotrophic strain is a dal/dat mutant.10ListeriaListeriaListeria. The recombinant strain of claim 9 , wherein said auxotrophic strain comprises an episomal expression vector comprising a ...

METHODS FOR CONTROLLING PROTEASE PRODUCTION

The present description is related to the field of protein production. It introduces novel host cells with low protease activity, a novel protease regulator, its use in expression systems and protein production, and a method of producing host cells for protein production. 1. A protein preparation comprising protein of interest produced in a host cell having at least one inactivated chromosomal gene wherein:the at least one inactivated chromosomal gene comprises a nucleic acid sequence encoding a polypeptide comprising a sequence having at least 90% sequence identity with the amino acids 402-533 of SEQ ID NO: 13;the at least one inactivated chromosomal gene is inactivated by disruption;and the host cell has reduced protease activity compared to the host cell without said inactivation.2. The protein preparation of claim 1 , wherein the protein preparation further comprises at least one further component selected from stabilizer claim 1 , preservative claim 1 , fragrant claim 1 , buffer claim 1 , salt and colorant.3. The protein preparation of claim 1 , wherein the protein preparation has an improved stability compared to a corresponding protein preparation produced in a host cell with an intact nucleic acid sequence encoding a polypeptide comprising a sequence having at least 90% sequence identity with the amino acids 402-533 of SEQ ID NO: 13.4. The protein preparation of claim 2 , wherein the protein preparation has an improved stability compared to a corresponding protein preparation produced in a host cell with an intact nucleic acid sequence encoding a polypeptide comprising a sequence having at least 90% sequence identity with the amino acids 402-533 of SEQ ID NO: 13.5. The protein preparation of claim 1 , wherein the protein preparation has a reduced amount of endogenous proteases compared to a corresponding protein preparation produced in a host cell with an intact nucleic acid sequence encoding a polypeptide comprising a sequence having at least 90% sequence ...

A Liquid Formulation of Alpha-Amylase

The present disclosure relates to liquid enzyme formulations containing one or more alpha-amylases for use in starch processing, wherein the pH of the enzyme formulation is about pH 6.0-8.0, and methods of use thereof. The present disclosure further relates to methods of making a liquid enzyme formulation containing one or more alpha-amylase having improved stability, comprising titrating the pH of the liquid enzyme formulation to a range of pH 6.0-8.0. 1. A liquid enzyme formulation comprising:(a) an alpha-amylase;(b) a buffering agent;(c) a stabilizer; and(d) a preservative,wherein the pH of the enzyme formulation is about pH 6.0-8.0.2. The liquid enzyme formulation of claim 1 , wherein the pH of the liquid enzyme formulation is about pH 6.3-6.7.3. The liquid enzyme formulation of claim 1 , wherein the stabilizer comprises sucrose claim 1 , sorbitol claim 1 , mannitol claim 1 , glycerol claim 1 , trehalose claim 1 , sodium chloride claim 1 , sodium sulfate claim 1 , or any combination thereof.4. The liquid enzyme formulation of claim 1 , wherein the buffering agent comprises: sodium citrate claim 1 , potassium citrate claim 1 , citric acid claim 1 , sodium acetate claim 1 , acetic acid claim 1 , sodium phosphate claim 1 , potassium phosphate claim 1 , or any combination thereof.5. The liquid enzyme formulation of claim 1 , wherein the alpha-amylase retains at least 90% of its activity at a temperature of 4-40° C.6. The liquid enzyme formulation of claim 1 , wherein the alpha-amylase retains at least 90% of its activity at a temperature of 25-30° C.7. The liquid enzyme formulation of claim 1 , wherein the alpha-amylase retains at least 90% of its activity for 1 year.8. The liquid enzyme formulation of claim 1 , wherein the alpha-amylase has a shelf life of at least 1 year.9. The liquid enzyme formulation of claim 8 , wherein the alpha-amylase has a shelf life of at least 1 year at 25° C.10. The liquid enzyme formulation of claim 1 , wherein the preservative ...

Methods For Purifying Insect Membrane-Bound Receptor Proteins From Recombinant Production Hosts

The invention is drawn to a method for purifying membrane-bound proteins expressed in recombinant insect cells using N-laurosarcosine. The invention is particularly suited for expressing cadherin-type receptors cloned from European Corn Borer, and expressed in Sf9 insect cells. The method is optionally adapted for use with 6-his tag proteins. 1. A method for purifying a membrane-bound protein from a recombinant insect cell which comprises:a) suspending the recombinant insect cells in a suitable lysis buffer containing N-laurylsarcosine for an effective period of time, and optionally sonicating;b) centrifuging the lysate from step a);c) dialyzing the supernatant from step b) against a suitable buffer containing 0.15% N-laurylsarcosine, and;d) isolating the membrane-bound protein using a chromatographic method.29. The method of claim 1 , wherein the recombinant insect cell is the Sf insect cell line.3. The method of claim 1 , wherein the suitable buffer is 50 mM Tris® claim 1 , 300 mM NaCl claim 1 , 0.1 mM dithiothreitol claim 1 , and 1.0% N-laurylsarcosine at pH4. The method of claim 1 , wherein the isolation is performed over a HiTrap® Q column or a HisTrap HP column. This application claims the benefit of priority under 35 U.S.C. §119(e) to U.S. Provisional Application Ser. No. 61/746,641, filed Dec. 28, 2012, the disclosure of which is incorporated herein by reference.This invention is in the field of biochemistry and molecular biology. In particular the invention relates to purification of membrane receptor proteins from recombinant host systems.Membrane receptor proteins located in epithelial cells of insect midguts are target sites for the action of crystalline protein toxins (cry toxins) produced by (Bt). Activated cry toxins bind to insect receptor proteins on insect midgut epithelial cells and cause toxicity and death to specific insect pests. The specific interactions between the receptor and toxin are key events in discerning the structure-activity ...

METHOD FOR PRODUCING A PROTEIN HYDROLYSATE EMPLOYING AN ASPERGILLUS FUMIGATUS TRIPEPTIDYL PEPTIDASE

The present invention relates to compositions and methods for the production of a hydrolysate comprising at least one endoprotease and a tripeptidyl peptidase capable of cleaving tripeptides from the N-terminus a peptide and/or proteins having one or more of lysine, arginine or glycine in the P1 position wherein said tripeptidyl peptidase is capable of being used at a temperature between 45° C. and 70° C. 1. A method for the production of a hydrolysate comprising: i) comprises the amino acid sequence SEQ ID No. 3, SEQ ID No. 4 or a functional fragment thereof;', 'ii) comprises an amino acid having at least 70% identity to SEQ ID No. 3 or SEQ ID No. 4;', 'iii) is encoded by a nucleotide sequence comprising the sequence SEQ ID No. 1 or SEQ ID No. 2;', 'iv) is encoded by a nucleotide sequence which has at least about 70% identity to SEQ ID No. 1 or SEQ ID No. 2;', 'v) is encoded by a nucleotide sequence which hybridises to SEQ ID No. 1 or SEQ ID No. 2 under medium stringency conditions; or', 'vi) is encoded by a nucleotide sequence which differs from SEQ ID No. 1 or SEQ ID No. 2 due to degeneracy of the genetic code;, 'a) admixing at least one protein or a portion thereof with a tripeptidyl peptidase whichb) incubating at a temperature between 45° C. and 70° C., andc) recovering the hydrolysate.2. A method according to wherein the temperature of the incubation is between 50° C. and 65° C.3. A method according to wherein the temperature of the incubation is between 55° C. and 65° C.4. A method according to claim 3 , wherein the method further comprises admixing the recovered hydrolysate with at least one feed or food ingredient.5. A method according to wherein the protein or portion thereof is further treated with an endoprotease.6. A method according to wherein the endoprotease and the tripeptidyl peptidase are added simultaneously.7. A method according to wherein the endoprotease and the tripeptidyl peptidase are added sequentially claim 6 , e.g. with the tripeptidyl ...

PROLINE TOLERANT TRIPEPTIDYL PEPTIDASES AND USES THEREOF

A method for the production of a hydrolysate comprising: (a) admixing at least one protein or a portion thereof with: (A) at least one endoprotease; and (B) (a′) at least one proline tolerant tripeptidyl peptidase or fermentate comprising a proline tolerant tripeptidyl peptidase predominantly having exopeptidase activity wherein said proline tolerant tripeptidyl peptidase is capable of cleaving tri-peptides from the N-terminus of peptides having: Proline at P1; and synthetic amino acids at P1; or (b′) at least one proline tolerant tripeptidyl peptidase having exopeptidase activity wherein said proline tolerant tripeptidyl peptidase is capable of cleaving tri-peptides from the N-terminus of peptides having: Proline at PV; and synthetic amino acids at PV; and (b) recovering the hydrolysate. The invention also relates to methods for producing a hydrolysate comprising the use of an endoprotease an exo-tripeptidyl peptidase of the S53 family and an aminopeptidase, to uses of a proline tolerant tripeptidyl peptidase, compositions, food and/or feed additive compositions comprising the same, as well as hydrolysates and uses of proline tolerant tripeptidyl peptidases. 1. A method for the production of a hydrolysate comprising: (A) at least one endoprotease; and', Proline at P1; and', 'an amino acid selected from alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, serine, threonine, tryptophan, tyrosine, valine or synthetic amino acids at P1; or', '(b′) at least one proline tolerant tripeptidyl peptidase having exopeptidase activity wherein said proline tolerant tripeptidyl peptidase is capable of cleaving tri-peptides from the N-terminus of peptides having:', 'Proline at P1′; and', 'an amino acid selected from alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, ...

METHOD FOR PREVENTING OR TREATING METABOLIC DISORDER

Provided is a method for preventing or treating a metabolic disorder, including administering to a subject a therapeutically effective amount of TRABID protein or a functionally related variant thereof, or a nucleic acid encoding TRABID protein or a functionally related variant thereof. Also provided is a method for reducing fat accumulation through TRABID-induced deubiquitination to promote autophagy activity and lipid metabolism. 1. A method for preventing or treating a metabolic disorder in a subject in need thereof , comprising administering to the subject a therapeutically effective amount of tumor-necrosis factor receptor-associated factor (TRAF)-binding protein domain (TRABID) protein or a functionally related variant thereof , or a nucleic acid encoding the TRABID protein or a functionally related variant thereof.2. The method according to claim 1 , wherein the TRABID protein comprises an amino acid sequence of SEQ ID NO:1 or SEQ ID NO:3 claim 1 , and the functionally related variant of the TRABID protein comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:1 or SEQ ID NO:3.3. The method according to claim 1 , wherein the nucleic acid encoding the TRABID protein comprises a nucleic acid sequence of SEQ ID NO:2 or SEQ ID NO:4 claim 1 , and the functionally related variant of the nucleic acid encoding the TRABID protein comprises a nucleic acid sequence having at least 80% sequence identity to SEQ ID NO:2 or SEQ ID NO:4.4. The method according to claim 1 , wherein the metabolic disorder is selected from the group consisting of pre-diabetes claim 1 , diabetes claim 1 , obesity claim 1 , diabetic dyslipidemia claim 1 , hyperlipidemia claim 1 , hypertriglyceridemia claim 1 , hyperfattyacidemia claim 1 , hypercholesterolemia claim 1 , fatty liver disease claim 1 , and any combination thereof.5. The method according to claim 4 , wherein the fatty liver disease is selected from the group consisting of non-alcoholic fatty liver disease ...

Method for reducing established metastatic tumor by pharmaceutical composition containing polypeptide

A method for reducing an established metastatic tumor, comprising administering an effective amount of a pharmaceutical composition to a subject in need, wherein the pharmaceutical composition comprises a polypeptide of a fibronectin-binding domain of dipeptidyl peptidase IV having a maltose-binding protein (MBP) fused at an N-terminal thereof, a cross-linking molecule, an active agent, and a pharmaceutically acceptable carrier.

ENZYME AND USES THEREOF

The present invention relates generally to methods and materials relating to newly characterised enzymes from , or variants thereof, capable degrading poly-γ-D-glutamic acid, for example as is present in the capsule of . Such depolymerases have utility as therapeutics. 1. A PGDA depolymerase polypeptide having a sequence which is at least about 60% , 70% , 80% or 85% identical to the amino acid sequence provided herein as SEQ ID NO: 2 , which comprises a His tag or which is a fusion protein of SEQ ID NO: 2 , or a fragment thereof with a heterologous fusion partner.2. (canceled)3. A PGDA depolymerase polypeptide comprising , consisting of , or consisting essentially of , the amino acid sequence provided herein as SEQ ID NO: 2 , or a fragment thereof , which lacks all or part of the signal peptide shown as amino acids 1 to 18 of SEQ ID NO: 2.49.-. (canceled)10. A vector comprising a nucleic acid having a sequence selected from the group consisting of:(a) a sequence encoding a PGDA depolymerase polypeptide having a sequence which is at least about 60%, 70%, 80% or 85% identical to the amino acid sequence provided herein as SEQ ID NO: 2;(b) a sequence which is complementary to, or which hybridises to, a sequence as defined in (a);(c) a fragment of a sequence as defined in (a) or (b); and(d) a sequence that is degenerate as a result of the genetic code to any one of (a) to (c); wherein the sequence is operably linked to a heterologous promoter.11. A host cell transformed with the vector of .12E. coli.. A host cell as claimed in wherein said cell is prokaryotic claim 11 , and which is preferably13. A host cell as claimed in which possesses mutations in thioredoxin reductase and\or glutathione reductase genes.14. A method of producing a host cell having enhanced PGDA depolymerase activity claim 10 , which method comprises introducing into said host a vector as claimed in .15. A process of producing a polypeptide claim 11 , the process comprising culturing a host cell as ...

Fusion proteins and methods for stimulating plant growth, protecting plants, and immobilizing bacillus spores on plants

The present invention is generally directed to fusion proteins containing a targeting sequence that targets the fusion protein to the exosporium of a Bacillus cereus family member. The invention also relates to recombinant Bacillus cereus family members expressing such fusion proteins and formulations containing the recombinant Bacillus cereus family members expressing the fusion proteins. Methods for stimulating plant growth, for protecting plants from pathogens, and for enhancing stress resistance in a plant by applying the recombinant Bacillus cereus family members or the formulations to plants or a plant growth medium are also described. The invention also relates to methods for immobilizing spores of a recombinant Bacillus cereus family member expressing a fusion protein on plants.

METHOD FOR PROPHYLAXIS AND/OR TREATMENT OF ErbB2 POSITIVE CANCERS

Provided are compositions and methods for prophylaxis and/or therapy of ErbB2-positive cancer. The compositions include pharmaceutical preparations that contain isolated or recombinant or modified peptidase D (PEPD) proteins. The methods include prophylaxis and/or therapy of ErbB2-positive cancer by administering a PEPD to an individual who has or is at risk for developing ErbB2-positive cancer. 1. A pharmaceutical preparation for use in prophylaxis and/or therapy of ErbB2-positive cancer comprising an isolated or recombinantly produced peptidase D (PEPD) and at least one pharmaceutically acceptable carrier.2. The pharmaceutical preparation of claim 1 , wherein the PEPD is a component of a fusion protein.3. The pharmaceutical preparation of claim 2 , wherein the fusion protein comprises the PEPD and a polyhistidine tag.4. The pharmaceutical composition of claim 1 , wherein the PEPD is coupled to a chemotherapeutic agent.5. The pharmaceutical preparation of claim 2 , wherein the fusion protein comprises the PEPD and an Fc region of an immunoglobulin.6. The pharmaceutical preparation of claim 1 , wherein the PEPD has less dipeptide hydrolysis activity as compared to a PEPD comprising the sequence of SEQ ID NO:1.7. The pharmaceutical preparation of claim 6 , wherein the PEPD comprises a mutation of glycine at position 278 of SEQ ID NO:1.8. The pharmaceutical preparation of claim 1 , further comprising a coagulation inhibitor.9. A product for use in prophylaxis and/or therapy of ErbB2-positive cancer comprising at least one sealed container which contains a pharmaceutical preparation comprising peptidase D (PEPD) claim 1 , wherein the product comprises printed material which provides an indication that the pharmaceutical preparation is for use in prophylaxis and/or therapy of ErbB2-positive cancer in an individual.10. The product of claim 9 , wherein the PEPD has less dipeptide hydrolysis activity as compared to a PEPD comprising the sequence of SEQ ID NO:1.11. The ...

FUSION PROTEINS AND METHODS FOR STIMULATING PLANT GROWTH, PROTECTING PLANTS, AND IMMOBILIZING BACILLUS SPORES ON PLANTS

The present invention is generally directed to fusion proteins containing a targeting sequence that targets the fusion protein to the exosporium of a family member. The invention also relates to recombinant family members expressing such fusion proteins and formulations containing the recombinant family members expressing the fusion proteins. Methods for stimulating plant growth, for protecting plants from pathogens, and for enhancing stress resistance in a plant by applying the recombinant family members or the formulations to plants or a plant growth medium are also described. The invention also relates to methods for immobilizing spores of a recombinant family member expressing a fusion protein on plants. 1239-. (canceled)240Bacillus cereus. A recombinant family member that expresses a fusion protein , wherein the fusion protein comprises:{'i': 'Bacillus cereus', 'a targeting sequence, exosporium protein, or exosporium protein fragment that targets the fusion protein to the exosporium of the recombinant family member; and'}an aminocyclopropane-1-carboxylic acid deaminase.241Bacillus cereus. The recombinant family member of claim 240 , wherein the fusion protein comprises:(a) a targeting sequence comprising an amino acid sequence having at least about 43% identity with amino acids 20-35 of SEQ ID NO: 1, wherein the identity with amino acids 25-35 is at least about 54%;(b) a targeting sequence comprising amino acids 1-35 of SEQ ID NO: 1;(c) a targeting sequence comprising amino acids 20-35 of SEQ ID NO: 1;(d) a targeting sequence comprising SEQ ID NO: 1;(e) an exosporium protein comprising an amino acid sequence having at least 85% identity with SEQ ID NO: 2;(f) a targeting sequence comprising amino acids 1-27 of SEQ ID NO: 3;(g) a targeting sequence comprising amino acids 12-27 of SEQ ID NO: 3;(h) a targeting sequence comprising SEQ ID NO: 3;(i) an exosporium protein comprising an amino acid sequence having at least 85% identity with SEQ ID NO: 4;(j) a targeting ...

Fusion Protease

This invention relates to novel bifunctional fusion proteases useful for manufacturing a mature protein from a fusion protein. More specifically the present invention relates to bifunctional fusion proteases comprising a picornaviral 3C protease and a Xaa-Pro-dipeptidyl aminopeptidase. 1. A bifunctional fusion enzyme comprising the catalytic domains of a picornaviral 3C protease and a XaaProDAP.2. The bifunctional fusion protease according to comprising a protein of the formula:{'br': None, 'X—Y—Z\u2003\u2003(I) or'}{'br': None, 'Z—Y—X\u2003\u2003(II)'}whereinX is a picornaviral 3C protease or a functional variant thereof;Y is an optional linker;Z is a Xaa-Pro-dipeptidyl aminopeptidase (XaaProDAP) or a functional variant thereof;wherein said fusion protease has substantially no self-cleavage activity able to deteriorate at least one of the two proteolytic activities.3. The bifunctional fusion protease according to comprising a protein of formula (I) claim 2 , wherein said picornaviral 3C protease or a functional variant thereof is in the N-terminal part of said bifunctional fusion protease.4. The bifunctional fusion protease according to claim 2 , wherein X is a human Rhinovirus 3C protease or a functional variant thereof.5. The bifunctional fusion protease according to claim 2 , wherein X comprises SEQ ID NO: 2 claim 2 , or a functional variant thereof.6. The bifunctional fusion protease according to claim 2 , wherein Z is an E.C. 3.4.14.11 enzyme or a functional variant thereof.7. The bifunctional fusion protease according to claim 6 , wherein Z is an enzyme from a lactic acid bacterium or a functional variant thereof.8. The bifunctional fusion protease according to claim 2 , wherein Z is SEQ ID NO: 1 or a functional variant thereof.9Streptococcus. The bifunctional fusion protease according to claim 2 , wherein Z is an enzyme from spp. or a functional variant thereof.10. The bifunctional fusion protease according to wherein Z is SEQ ID NO: 24 or a functional ...

FUSION PROTEINS AND METHODS FOR STIMULATING PLANT GROWTH, PROTECTING PLANTS FROM PATHOGENS, AND IMMOBILIZING BACILLUS SPORES ON PLANT ROOTS

The present invention is generally directed to fusion proteins containing a targeting sequence that targets the fusion protein to the exosporium of a family member. The invention also relates to recombinant family members expressing such fusion proteins, formulations containing the recombinant family members expressing the fusion proteins. Methods for stimulating plant growth and for protecting plants from pathogens by applying the recombinant family members or the formulations to plants or a plant growth medium are also described. The invention also relates to methods for immobilizing spores of a recombinant family member expressing a fusion protein on plant roots. 1152-. (canceled)153Bacillus cereus. A recombinant family member that expresses a fusion protein , wherein the fusion protein comprises:{'i': 'Bacillus cereus', 'a targeting sequence or exosporium protein that is capable of targeting the fusion protein to the exosporium of the recombinant family member; and'}at least one plant growth stimulating protein or peptide, wherein the plant growth stimulating protein or peptide comprises an endoglucanase, a chitosanase, a protease, a glycoside hydrolase, a phosphatase, a nitrogenase, a nuclease, a glucosidase, a glucanase, a flagellin protein, or a flagellin peptide.154Bacillus cereus. A recombinant family member of claim 153 , wherein the fusion protein comprises a targeting sequence comprising an amino acid sequence having at least about 43% identity with amino acids 20-35 of SEQ ID NO: 1 claim 153 , wherein the identity with amino acids 25-35 is at least about 54%.155Bacillus cereus. A recombinant family member of claim 153 , wherein the fusion protein comprises a targeting sequence comprising an amino acid sequence having at least about 50% identity with amino acids 20-35 of SEQ ID NO: 1 claim 153 , wherein the identity with amino acids 25-35 is at least about 63%.156Bacillus cereus. A recombinant family member of claim 153 , wherein the fusion protein ...

Methods and compositions for the treatment of amyloidosis

Methods and compositions for the treatment or prevention of amyloidosis are provided. In some embodiments, the methods comprise administering to the subject a therapeutically effective amount of at least one catabolic enzyme or a biologically active fragment thereof. Such methods and compositions may be employed to reduce, prevent, degrade and/or eliminate amyloid formation in the lysosome and/or extracellularly.

MODIFIED ESCHERICHIA COLI STRAIN NISSLE AND TREATMENT OF GASTROINTESTINAL DISORDER

The invention relates to the field of modified strain Nissle 1917 (EcN) and its use for treating gastro-in-testinal disorders. The invention is based on the study of the mechanisms implicated in the probiotic properties of the strain Nissle 1917 (EcN). This study has allowed the inventors to decouple the probiotic activity of EcN from its genotoxic activity by demonstrating that EcN ClbP protein, the enzyme that activates the genotoxin colibactin, is also required for the siderophore-microcins activity of probiotic EcN, but interestingly, not its enzymatic domain that cleaves precolibactin to form active colibactin. Furthermore, inventors demonstrate in an in vivo animal model infected by a bacterial pathogen that administration of an EcN modified strain with clbP gene encoding ClbP protein inactive for the peptidase domain, is non-genotoxic (do not produce colibactin) but keeps the bacterial antagonist activity, and reduces colonization and virulence of the pathogen by maintaining the siderophore-microcin production. Thus this study opens the way to safe use of EcN and accordingly the present invention provides an strain Nissle 1917 (EcN) bacterium carrying a gene encoding ClbP protein which is inactive for the peptidase domain, and its use as a drug and more particularly for use in the treatment of gastro-intestinal disease. 1Escherichia coli. An strain Nissle 1917 (EcN) bacterium carrying a gene encoding a ClbP protein comprising an inactive peptidase domain ,wherein said EcN bacterium (i) has antibacterial activity; and ii) is devoid of the capacity to activate the genotoxin colibactin.2. The EcN bacterium according to wherein the ClbP protein comprising an inactive peptidase domain claim 1 , is selected from the group consisting of:ClbP protein mutated at position S95,ClbP protein mutated at position K98,ClbP protein mutated at position Y186, andClbP protein without peptidase domain.3. The EcN bacterium according to claim 2 , wherein the ClbP protein is mutated ...

VECTOR-BASED MUTAGENESIS SYSTEM

Strategies, reagents, methods, and systems for modulating the mutation rate in cells are provided herein. The strategies, reagents, methods, and systems are broadly applicable for the modulation of mutation rates in cells where high mutation rates and/or control over a broad range of mutation rates is desired, for example, in the context of diversifying a nucleic acid sequence or a plurality of such sequences within a population of cells, for the generation of diversified nucleic acid libraries, and for directed evolution of nucleic acids and encoded products. 1125-. (canceled)126. An expression construct for modulating the mutation rate of nucleic acids in a bacterial cell , the construct comprising:(i) a drift promoter; and(ii) a nucleic acid sequence encoding one or more gene products selected from a gene product that disrupts a proofreading pathway, a translesion synthesis pathway, a methyl-directed mismatch repair pathway, a base excision repair pathway, or a base selection pathway of the bacterial cell, or any combination thereof,wherein the nucleic acid sequence encoding the one or more gene products is under the control of the drift promoter.127. The expression construct of claim 126 , wherein the gene product that disrupts a proofreading pathway is a dnaQ926 claim 126 , BRM1 claim 126 , BR11 claim 126 , BR1 claim 126 , BR6 claim 126 , or BR13 gene product.128. The expression construct of claim 126 , wherein the gene product that disrupts a translesion synthesis pathway is an umuD′ claim 126 , umuC claim 126 , recA claim 126 , dinB claim 126 , or polB gene product.129. The expression construct of claim 126 , wherein the gene product that disrupts a methyl-directed mismatch repair pathway is a mutS claim 126 , mutL claim 126 , mutH claim 126 , dam claim 126 , or seqA gene product.130. The expression construct of claim 126 , wherein the gene product that disrupts a base excision repair pathway is a ugi claim 126 , AID claim 126 , APOBEC claim 126 , CDA claim ...

VACCINIA VIRUS FOR GENE-DIRECTED ENZYME PRODRUG THERAPY

Methods of making vaccinia viruses for gene-directed prodrug therapy are disclosed and their use in the treatment of disease is provided. In particular, the anti-tumor effects of vaccinia viruses that are modified to express a prodrug-activating enzyme are disclosed. 121.-. (canceled)22. A vaccinia virus which is capable of expressing a glutamate carboxypeptidase within a cell.23. The vaccinia virus according to claim 22 , wherein the glutamate carboxypeptidase is CPG2 or a mutant claim 22 , variant or homologue thereof.24. The vaccinia virus according to claim 23 , wherein the glutamate carboxypeptidase comprises an amino acid sequence which has at least 80% identity to SEQ ID NO: 2.25. The vaccinia virus according to claim 23 , wherein the vaccinia virus comprises a CPG2 expression cassette insert in a vaccinia virus virulence gene.26. The vaccinia virus according to claim 25 , wherein the vaccinia virus virulence gene is vaccinia thymidine kinase (vTK).27. A two component system for gene directed enzyme prodrug therapy which comprises: (a) a vaccinia virus according to ; and (b) a prodrug which can be converted into an active drug by a glutamate carboxypeptidase.29. The two component system according to claim 28 , wherein the linker comprises a functional group selected from: carbonyl claim 28 , carbamate claim 28 , urea claim 28 , and equivalents.30. The two component system according to claim 28 , wherein the cytotoxic agent comprises a nitrogen mustard or a nitrogen mustard analogue.31. The two component system according to claim 30 , wherein the prodrug is N-(4-[bis(2-iodoethyl)amino]phenoxycarbonyl)-L-glutamic acid or a salt thereof.32. A method of killing a neoplastic cell claim 22 , the method comprising treating the neoplastic cell with: a vaccinia virus according to and a prodrug which is capable of being converted into an active form by a glutamate carboxypeptidase.33. The method of killing a neoplastic cell according to claim 32 , wherein the prodrug ...

CLEANING AGENT CONTAINING PROTEASES

A cleaning agent is proposed that includes a first and a second protease, wherein a) the first protease comprises an amino acid sequence which is at least 80% identical to the amino acid sequence specified in SEQ ID NO. 1 over the entire length and has at least one amino acid substitution at one of the positions 9, 15, 66, 212, and 239 using the numbering according to SEQ ID NO. 1, and b) the second protease comprises an amino acid sequence which is at least 80% identical to the amino acid sequence specified in SEQ ID NO. 2 over the entire length and has at least one amino acid substitution at one of the positions 32, 33, 48-54, 58-62, 94-107, 116, 123-133, 150, 152-156, 158-161, 164, 169, 175-186, 197, 198, and 203-216 using the numbering according to SEQ ID NO. 2. 1. A cleaning agent comprising at least one first protease and one second protease , wherein:a. the first protease comprises an amino acid sequence which is at least about 80% identical to the amino acid sequence specified in SEQ ID NO. 1 over the entire length and has at least one amino acid substitution at one of the positions 9, 15, 66, 212, and 239 using the numbering according to SEQ ID NO. 1, andb. the second protease comprises an amino acid sequence which is at least about 80% identical to the amino acid sequence specified in SEQ ID NO. 2 over the entire length and has at least one amino acid substitution at one of the positions 32, 33, 48-54, 58-62, 94-107, 116, 123-133, 150, 152-156, 158-161, 164, 169, 175-186, 197, 198, and 203-216 using the numbering according to SEQ ID NO. 2.2. The cleaning agent according to claim 1 , wherein the first protease has at least one amino acid substitution selected from the group consisting of S9R claim 1 , A15T claim 1 , V66A claim 1 , N212D claim 1 , and Q239R claim 1 , using the numbering according to SEQ ID NO. 1.3. The cleaning agent according to claim 1 , wherein the first protease has an amino acid sequence according to SEQ ID NO. 3.4. The cleaning agent ...

ENDOGLUCANASE VARIANTS

The present invention relates to variant endoglucanases having improved thermoactivity, improved thermostability, and improved viscosity reduction activity over wild-type endoglucanase. 154.-. (canceled)55. A polynucleotide encoding a variant endoglucanase polypeptide having endoglucanase activity , wherein the polypeptide comprises:(a) an amino acid sequence having at least 80% sequence identity to a wild-type endoglucanase 2 (SEQ ID NO:1); and(b) an amino acid substitution of at least one of the amino acid residues L71, D118, N202, 5242, or M250, wherein the positions are numbered with reference to SEQ ID NO:1.56. The polynucleotide of claim 55 , wherein the variant comprises an amino acid substitution of each of the amino acid residues L71 claim 55 , D118 claim 55 , N202 claim 55 , 5242 claim 55 , and M250; and has an amino acid substitution of one or more of the amino acid residues selected from K70 claim 55 , K72 claim 55 , S76 claim 55 , E82 claim 55 , E85 claim 55 , N87 claim 55 , L91 claim 55 , K94 claim 55 , H95 claim 55 , Q128 claim 55 , 5131 claim 55 , D134 claim 55 , E135 claim 55 , G167 claim 55 , N174 claim 55 , R177 claim 55 , Q186 claim 55 , N190 claim 55 , T203 claim 55 , D205 claim 55 , N211 claim 55 , G224 claim 55 , T226 claim 55 , V232 claim 55 , A236 claim 55 , T245 claim 55 , T248 claim 55 , T254 claim 55 , Q257 claim 55 , 5270 claim 55 , A277 claim 55 , S282 claim 55 , A286 claim 55 , Q287 claim 55 , V289 claim 55 , V290 claim 55 , A298 claim 55 , N299 claim 55 , L302 claim 55 , A312 claim 55 , L338 claim 55 , Y351 claim 55 , G360 claim 55 , N363 claim 55 , and S366 claim 55 , wherein the residues are numbered with reference to SEQ ID NO:1.57. The polynucleotide of claim 55 , wherein the variant comprises the amino acid substitutions L71F claim 55 , D118P claim 55 , N202H claim 55 , S242T claim 55 , and M250L; and has one or more amino acid substitutions selected from K70R claim 55 , K72E/H claim 55 , S76A/M/V claim 55 , E82K claim 55 , E85A/ ...

FUSION PROTEINS AND METHODS FOR STIMULATING PLANT GROWTH, PROTECTING PLANTS FROM PATHOGENS, AND IMMOBILIZING BACILLUS SPORES ON PLANT ROOTS

The present invention is generally directed to fusion proteins containing a targeting sequence that targets the fusion protein to the exosporium of a family member. The invention also relates to recombinant family members expressing such fusion proteins, formulations containing the recombinant family members expressing the fusion proteins. Methods for stimulating plant growth and for protecting plants from pathogens by applying the recombinant family members or the formulations to plants or a plant growth medium are also described. The invention also relates to methods for immobilizing spores of a recombinant family member expressing a fusion protein on plant roots. 1152-. (canceled)153Bacillus cereusBacillus cereusBacillus cereus. A plant seed coated with a recombinant family member , wherein the recombinant family member expresses a fusion protein and the fusion protein comprises at least one protein or peptide of interest and a targeting sequence or exosporium protein that targets the fusion protein to the exosporium of the recombinant family member.154Bacillus cereus. A plant seed of claim 153 , wherein the plant seed is coated with a seed coating formulation comprising the recombinant family member and an agriculturally acceptable carrier.155. A plant seed of claim 153 , wherein the fusion protein comprises a targeting sequence comprising an amino acid sequence having at least about 43% identity with amino acids 20-35 of SEQ ID NO: 1 claim 153 , wherein the identity with amino acids 25-35 is at least about 54%.156. A plant seed of claim 153 , wherein the targeting sequence comprises an amino acid sequence having at least about 50% identity with amino acids 20-35 of SEQ ID NO: 1 claim 153 , wherein the identity with amino acids 25-35 is at least about 63%.157. A plant seed of claim 153 , wherein the targeting sequence comprises an amino sequence having at least about 62% identity with amino acids 20-35 of SEQ ID NO: 1 claim 153 , wherein the identity with amino ...

UBIQUITIN CHAIN ANALYSIS

There is described a method for analyzing ubiquitin polymers using linkage-specific deubiquitinase enzymes. Novel specificities of deubiquitinase enzymes are also provided. 1. A method for determining the linkage type in a polyubiquitin chain , comprising the steps of:(a) contacting an ubiquitin polymer with a first linkage-specific deubiquitinase under conditions in which the enzyme catalyses the disassociation of one or more, but less than seven, ubiquitin linkages;(b) analysing the product of the catalysis for the presence of lower molecular weight fractions of the ubiquitin polymer; and(c) repeating steps (a) and (b) in the presence of a second, different linkage-specific deubiquitinase.2. The method according to claim 1 , wherein the deubiquitinase is an ovarian tumour protease (OTU) family deubiquitinase (DUB).3. The method according to claim 1 , wherein the deubiquitinase catalyses the dissociation of three claim 1 , two or one linkage types.4. The method according to claim 1 , wherein the product of the catalysis reaction is analysed by gel electrophoresis.5. The method according to claim 1 , wherein the lower molecular weight fractions comprise ubiquitin monomers and/or polymers.6. The method according to claim 1 , wherein the second linkage-specific deubiquitinase has a linkage specificity which overlaps with that of the first linkage-specific deubiquitinase.7. The method according to claim 1 , wherein the second linkage-specific deubiquitinase has a linkage specificity which does not overlap with that of the first linkage-specific deubiquitinase.8. The method according to claim 1 , wherein ovarian tumour protease family deubiquitinase 1 (OTUD1) is used to cleave K63 linkages.9. The method according to claim 1 , wherein ovarian tumour protease family deubiquitinase 4 (OTUD4) is used to cleave K48 linkages.10. The method according to claim 1 , wherein ovarian tumour protease family deubiquitinase 3 (OTUD3) is used to cleave K6 and/or K11 linkages.11. A ...

METHOD FOR HARVESTING ORGANIC COMPOUNDS FROM GENETICALLY MODIFIED ORGANISMS

Disclosed herein are embodiments for a novel method of producing an organic compound, including harvesting at least one organic compound from an organism or cell line genetically engineered with a gene for at least one proton-pump protein. 1. A method of producing an organic compound comprising ,harvesting at least one organic compound from an organism or cell line genetically engineered with a gene for at least one proton-pump protein.2. The method of claim 1 , wherein the at least one proton-pump protein is a light-driven proton-pump protein.3. The method of claim 1 , wherein the at least one proton-pump protein is an archaerhodopsin claim 1 , bacteriorhodopsin claim 1 , opsin claim 1 , proteorhodopsin claim 1 , rhodopsin claim 1 , xanthorhodopsin claim 1 , homologs thereof or combinations thereof.4. The method of claim 3 , wherein the at least one proton-pump protein absorbs light between about 100 nm and about 1 μm.5. The method of claim 4 , wherein the at least one proton-pump protein absorbs light between about 300 nm and about 750 nm.6. The method of claim 5 , wherein the at least one proton-pump protein absorbs light between about 450 nm and about 650 nm.7. The method of claim 6 , wherein the at least one proton-pump protein absorbs light between about 450 nm and about 550 nm.8. The method of claim 6 , wherein the at least one proton-pump protein absorbs light between about 550 nm and about 600 nm.9. The method of claim 1 , wherein the at least one organic compound is a biologic or a biofuel10. The method of claim 9 , wherein the biologic is selected from the group consisting of allergenics claim 9 , antibodies claim 9 , blood products or derivative thereof claim 9 , enzymes claim 9 , growth factors claim 9 , hormones claim 9 , immunomodulators claim 9 , interferons claim 9 , interleukins claim 9 , polypeptides claim 9 , proteins claim 9 , serum claim 9 , tissues claim 9 , toxins and vaccines.11. The method of claim 10 , wherein the antibody is selected from ...

PROTEOGLYCAN IRREGULARITIES IN ABNORMAL FIBROBLASTS AND THERAPIES BASED THEREFROM

Provided herein are methods to identify agents or compounds that specifically modulate the oligomerization and/or functional activities of receptor protein tyrosine phosphatase sigma (RPTPσ) in an abnormal fibroblast cell and therapies based therefrom. 1. A method of treating arthritis in a subject , the method comprising:obtaining a biological sample comprising synovial cells, synovial-like cells and/or synovial fluid from the joint of the subject;contacting the biological sample with a test agent that inhibits clustering and/or promotes PTP activity of receptor protein tyrosine phosphatase sigma (RPTPσ); anddetermining whether (i) there is a change in the clustering and/or biological activity of RPTPσ, or (ii) whether the agent binds to a ligand of the RPTPσ ectodomain;wherein if there is an inhibition of RPTPσ clustering and/or increase in PTP activity, or binding of the test agent to the ligand of the RPTPσ ectodomain the subject is treated with an agent that inhibits RPTPσ clustering or promotes PTP activity and/or RPTPσ biological activity.2. The method of claim 1 , wherein the agent is a soluble extracellular domain of RPTPσ.3. The method of claim 1 , wherein the agent is an RPTPσ Ig1&2 polypeptide.4. The method of claim 1 , wherein the agent is an antibody that specifically interacts with the RPTPσ ectodomain and inhibits clustering.5. The method of claim 1 , further comprising measuring the level of syndecan-4.6. A method of screening a subject having or at risk of having arthritis claim 1 , the method comprising:obtaining fibroblast-like synoviocytes (FLS) from a subject;contacting the FLS cells with an agent that inhibits clustering and/or promotes PTP activity of receptor protein tyrosine phosphatase sigma (RPTPσ), anddetermining whether (i) there is a change in the clustering and/or biological activity of RPTPσ on the FLS cells, or (ii) whether the agent binds to a ligand of the RPTPσ ectodomain on the FLS cells,wherein if there is an inhibition of RPTP ...

Anti-vegf protein compositions and methods for producing the same

The present disclosure pertains to compositions comprising aflibercept and methods for producing such compositions in chemically defined media and using chromatography to reduce amounts of certain aflibercept variants.

Consensus Prostate Antigens, Nucleic Acid Molecule Encoding The Same And Vaccine And Uses Comprising The Same

Provided herein are consensus amino acid sequences of prostate antigens that are capable of breaking tolerance in a targeted species, including PSA, PSMA, STEAP and PSCA antigens. Also provided are nucleic acid sequences that encode one or more consensus amino acid sequences of prostate antigens PSA, PSMA, STEAP and PSCA, as well as genetic constructs/vectors and vaccines expressing the sequences. Also provided herein are methods for generating an autoimmune response against prostate cancer cells by administering one or more of the vaccines, proteins, and/or nucleic acid sequences that are provided. 1. A nucleic acid molecule comprising a coding sequence encoding one or more proteins selected from the group comprising:a) SEQ ID NO:2, a protein that is 98% homologous to SEQ ID NO:2, provided amino acids 69, 78, 80, 82, 102, 110, 137, 139, 165, 189, 203, 220, 232 and 248 of SEQ ID NO:2 are conserved, or an immunogenic fragment of SEQ ID NO:2 comprising amino acids corresponding to at least 256 amino acid residues of SEQ ID NO:2 provided amino acids 69, 78, 80, 82, 102, 110, 137, 139, 165, 189, 203, 220, 232 and 248 of SEQ ID NO:2 are conserved;b) SEQ ID NO:4, a protein that is 98% homologous to SEQ ID NO:4, provided amino acids 21, 86, 127, 129, 154, 156, 182, 195, 206, 218, 220, 237, 249, 255, 265, 271 or 275 of SEQ ID NO:4 are conserved, or an immunogenic fragment of SEQ ID NO:4 comprising amino acids corresponding to at least 274 amino acid residues of SEQ ID NO:4, provided amino acids 21, 86, 127, 129, 154, 156, 182, 195, 206, 218, 220, 237, 249, 255, 265, 271 or 275 of SEQ ID NO:4 are conserved;c) SEQ ID NO:6, a protein that is 98% homologous to SEQ ID NO:6, provided amino acids 14, 15, 32, 47, 58, 79, 111, 157, 223, 320, 350, 475, 499, 569, 613, 624, 653, 660, 663, 733 and 734 of SEQ ID NO:6 are conserved, or an immunogenic fragment of SEQ ID NO:6 comprising amino acids corresponding to at least 735 amino acid residues of SEQ ID NO:6, provided amino acids 14, 15 ...

PERFORMANCE-ENHANCED AND TEMPERATURE-RESISTANT PROTEASE VARIANTS

Proteases that comprise an amino acid sequence that is at least 70% identical to the amino acid sequence indicated in SEQ ID NO. 1 over its entire length and that comprise, in the count in accordance with SEQ ID NO. 1, the amino acid substitution R99E or R99D in combination with at least two further amino acid substitutions that are selected from the group consisting of S3T, V4I, and V199I, display very good cleaning performance in particular on blood-containing stains, as well as very good temperature stability. 1. A protease comprising an amino acid sequence that is at least 70% identical to the amino acid sequence indicated in SEQ ID NO. 1 over its entire length and comprises , in the count in accordance with SEQ ID NO. 1 , the amino acid substitution R99E or R99D in combination with at least two further amino acid substitutions that are selected from the group consisting of S3T , V4I , and V199I.2. A protease , selected from the group consisting of proteases obtainable from:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, '(a.) protease according to as a starting molecule by single or multiple conservative amino acid substitution, wherein the protease comprises, in the count in accordance with SEQ ID NO. 1, the amino acid substitution R99E or R99D in combination with at least two further amino acid substitutions that are selected from the group consisting of S3T, V4I, and V199I;'}{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, '(b.) protease according to as a starting molecule by fragmentation, deletion mutagenesis, insertion mutagenesis, or substitution mutagenesis, and comprises an amino acid sequence that corresponds to the starting molecule over a length of at least 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 265, or 266 continuously connected amino acids, wherein the amino acid substitution R99E or R99D contained in the starting molecule, in combination with at least two further amino acid ...

PSMA BINDING LIGAND-LINKER CONJUGATES AND METHODS FOR USING

Described herein are prostate specific membrane antigen (PSMA) binding conjugates that are useful for delivering therapeutic, diagnostic and imaging agents. Also described herein are pharmaceutical compositions containing them and methods of using the conjugates and compositions. Also described are processes for manufacture of the conjugates and the compositions containing them. 135-. (canceled)36. A conjugate or a pharmaceutically acceptable salt thereof , the conjugate or the pharmaceutically acceptable salt thereof comprising a ligand of PSMA (B) , a linker (L) , and a drug (D) , wherein the ligand a carbon-sulfur double bond , a phosphorus-sulfur double bond , a phosphorus-sulfur single bond , a thioester , or a combination thereof wherein the linker is covalently bound to the drug via a hydrazide linkage , wherein the linker is covalently bound to the ligand wherein the linker comprises a chain of at least seven atoms , and wherein the linker comprises a disulfide linkage.39. The conjugate or the pharmaceutically acceptable salt thereof of claim 37 , wherein q is 1.40. The conjugate or the pharmaceutically acceptable salt thereof of claim 37 , wherein Y is independently selected in each instance from —CRR— claim 37 , and —NR—.41. The conjugate or the pharmaceutically acceptable salt thereof of claim 36 , wherein the linker comprises a chain of at least 14 atoms.42. The conjugate or the pharmaceutically acceptable salt thereof of claim 36 , wherein the linker comprises a peptide.43. The conjugate or the pharmaceutically acceptable salt thereof of claim 36 , wherein the linker comprises one or more phenylalanine residues claim 36 , each of which is independently optionally substituted.44. The conjugate or the pharmaceutically acceptable salt thereof of claim 36 , wherein the linker comprises phenylalanyl-phenylalanyl claim 36 , each of which is independently optionally substituted.47. The conjugate or the pharmaceutically acceptable salt thereof of claim 36 , ...

METHOD FOR PRODUCING USEFUL SUBSTANCE

The present disclosure concerns a method for producing peptides such as glutathione and a microorganism that can be used for such method. One or more embodiments of the first aspect of the present disclosure concern a method for producing peptides such as glutathione comprising culturing a prokaryotic microbial strain in which the expression levels of one or more genes selected from among the gshA gene, the gshB gene, and the gshF gene are enhanced, compared with the expression levels thereof in the wild-type strain thereof in a medium in which the total concentration of cysteine and cystine is 0.5 g/l or lower. The second aspect of the present disclosure concerns a microorganism comprising disruptions of the γ-glutamyltransferase gene and the glutathione reductase gene and exhibiting the enhanced expression levels of the gshA gene and the gshB or gshF gene. 1. A method for producing γ-glutamylcysteine , bis-γ-glutamylcystine , γ-glutamylcystine , reduced glutathione , and/or oxidized glutathione , comprisingculturing a gram-negative bacterium in a medium, wherein the total concentration of cysteine and cystine in the medium is 0.5 g/l or lower before inoculation of the gram-negative bacterium, thereby increasing the expression levels of one or more genes selected from genes encoding glutamate-cysteine ligase, glutathione synthetase, and bifunctional glutathione synthetase, when compared with expression levels in a wild-type strain thereof, wherein the gram negative bacterium is capable of overproducing γ-glutamylcysteine, bis-γ-glutamylcystine, γ-glutamylcystine, reduced glutathione, and/or oxidized glutathione by induced expression of the one or more genes when cultured in medium.2. The method according to claim 1 , wherein the gram-negative bacterium carries one or more genes selected from genes encoding glutamate-cysteine ligase claim 1 , glutathione synthetase claim 1 , and bifunctional glutathione synthetase claim 1 , operably linked to an inducible promoter ...

BMP PEPTIDES & METHODS OF USE

The invention relates to truncated growth factors and variants thereof. The invention also relates to methods of making and using the truncated growth factors. The invention further relates to compositions including a protease and a growth factor comprising a bone morphogenic protein (BMP) or a variant thereof. The invention also relates to methods of using the composition. 1. A method of promoting differentiation of cells into osteoblasts or chondrocytes , the method comprising treating the cells with a peptide selected from the group consisting ofa) a peptide of 115 residues or less, said peptide comprising an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 19,b) a peptide of 115 residues or less, said peptide comprising an amino acid sequence of SEQ ID NO: 19, andc) a peptide with an amino acid sequence consisting of the amino acid sequence of SEQ ID NO: 19,wherein each of peptides (a)-(c) has osteoinductive or chondroinductive activity.2. The method of claim 1 , wherein the cells are progenitor cells or adult stem cells.3. The method of claim 2 , wherein the progenitor cells or the adult stem cells are derived from placenta claim 2 , bone marrow claim 2 , adipose tissue claim 2 , blood vessel claim 2 , amniotic fluid claim 2 , synovial fluid claim 2 , synovial membrane claim 2 , pericardium claim 2 , periosteum claim 2 , dura claim 2 , peripheral blood claim 2 , umbilical blood claim 2 , menstrual blood claim 2 , baby teeth claim 2 , nucleus pulposus claim 2 , brain claim 2 , skin claim 2 , hair follicle claim 2 , intestinal crypt claim 2 , neural tissue claim 2 , or muscle.4. The method of claim 1 , wherein the cells are induced pluripotent stem cells.5. A method of promoting osteogenesis of cells in a tissue claim 1 , the method comprising treating the tissue with a peptide selected from the group consisting ofa) a peptide of 115 residues or less, said peptide comprising an amino acid sequence at least 95% identical to the ...

MICROORGANISMS PROGRAMMED TO PRODUCE IMMUNE MODULATORS AND ANTI-CANCER THERAPEUTICS IN TUMOR CELLS

Genetically programmed microorganisms, such as bacteria or virus, pharmaceutical compositions thereof, and methods of modulating and treating cancers are disclosed. 1. A modified microorganism capable of producing at least one immune initiator and at least one immune sustainer.2. The modified microorganism of claim 1 , wherein the immune initiator is capable of enhancing oncolysis claim 1 , activating antigen presenting cells (APCs) claim 1 , and/or priming and activating T cells.3. The modified microorganism of or claim 1 , wherein the immune initiator is a therapeutic molecule encoded by at least one gene; a therapeutic molecule produced by an enzyme encoded by at least one gene; at least one enzyme of a biosynthetic or catabolic pathway encoded by at least one gene; at least one therapeutic molecule produced by at least one enzyme of a biosynthetic or catabolic pathway encoded by at least one gene; or a nucleic acid molecule that mediates RNA interference claim 1 , microRNA response or inhibition claim 1 , TLR response claim 1 , antisense gene regulation claim 1 , target protein binding claim 1 , or gene editing.4. The modified microorganism of any one of - claim 1 , wherein the immune initiator is a cytokine claim 1 , a chemokine claim 1 , a single chain antibody claim 1 , a ligand claim 1 , a metabolic converter claim 1 , a T cell co-stimulatory receptor claim 1 , a T cell co-stimulatory receptor ligand claim 1 , an engineered chemotherapy claim 1 , or a lytic peptide.5. The modified microorganism of any one of - claim 1 , wherein the immune initiator is a STING agonist claim 1 , arginine claim 1 , 5-FU claim 1 , TNFα claim 1 , IFNγ claim 1 , IFNβ1 claim 1 , agonistic anti-CD40 antibody claim 1 , CD40L claim 1 , SIRPα claim 1 , GMCSF claim 1 , agonistic anti-OXO40 antibody claim 1 , OXO40L claim 1 , agonistic anti-4-1BB antibody claim 1 , 4-1BBL claim 1 , agonistic anti-GITR antibody claim 1 , GITRL claim 1 , anti-PD1 antibody claim 1 , anti-PDL1 antibody claim ...

COMPOSITION AND VACCINE FOR TREATING PROSTATE CANCER

The present invention relates to a composition comprising at least one mRNA encoding a combination of antigens capable of eliciting an (adaptive) immune response in a mammal, wherein the antigens are selected from the group consisting of PSA (Prostate-Specific Antigen), PSMA (Prostate-Specific Membrane Antigen), PSCA (Prostate Stem Cell Antigen), STEAP (Six Transmembrane Epithelial Antigen of the Prostate), MUC1 (Mucin 1) and PAP (Prostatic acid phosphatase). The invention furthermore relates to a vaccine comprising at least one mRNA encoding such a combination of antigens and to the use of said composition (for the preparation of a vaccine) and/or of the vaccine for eliciting an (adaptive) immune response for the treatment of prostate cancer (PCa), preferably of prostate adenocarcinoma, locally limited, locally advanced, metastatic, castration-resistant (hormone-refractory), metastatic castration-resistant and non-metastatic castration-resistant prostate cancers, and diseases or disorders related thereto. Finally, the invention relates to kits, particularly to kits of parts, containing the composition and/or the vaccine. 2. The composition according to claim 1 , wherein each of the antigens or fragments thereof is encoded by a separate mRNA.3. The composition according to claim 1 , wherein STEAP claim 1 , PSA claim 1 , PSMA claim 1 , PSCA claim 1 , PAP and MUC1 or fragments thereof are encoded by one mRNA.4. The composition according to claim 1 , wherein the antigens or fragments thereof are encoded by at least one bicistronic and/or multicistronic mRNA.5. The composition according to any of to claim 1 , wherein at least one mRNA claim 1 , preferably at least two mRNAs claim 1 , more preferably at least three mRNAs claim 1 , even more preferably at least four mRNAs claim 1 , even more preferably at least five mRNAs claim 1 , or even more preferably at least six mRNAs claim 1 , each comprise at least one coding sequence selected from RNA sequences being identical or ...

Compositions and methods for using fixed biological samples in partition-based assays

The present disclosure provides compositions and methods for using fixed biological samples in partition-based assays. In at least one embodiment, the disclosure provides a composition comprising a fixed biological sample and an un-fixing agent contained in a partition, such as a discrete droplet. In some embodiments, the disclosure provides un-fixing agent compounds capable of catalytically cleaving crosslinks in fixed biological samples, particularly crosslinked nucleic acids, such as RNA.

COMPOSITIONS AND METHODS FOR ADOPTIVE CELL THERAPY FOR CANCER

Provided herein are compositions and methods for adoptive cell therapy comprising engineered immune cells that express a tumor antigen-targeted chimeric antigen receptor and a prodrug converting enzyme. 1. An engineered immune cell comprising:(a) a first prodrug converting enzyme and/or a nucleic acid encoding the first prodrug converting enzyme, wherein the first prodrug converting enzyme is expressed in the cytoplasm of the immune cell and is selected from among a carboxypeptidase, a β-lactamase, a glucosidase, a nitroreductase, and carboxypeptidase A; and(b) a receptor that binds to a target antigen and/or nucleic acid encoding the receptor.2. The engineered immune cell of claim 1 , further comprising a second prodrug converting enzyme and/or a nucleic acid encoding the second prodrug converting enzyme claim 1 , wherein the second prodrug converting enzyme is expressed on the surface of the immune cell or is secreted.3. An engineered immune cell comprising:(a) a first prodrug converting enzyme and/or a nucleic acid encoding the first prodrug converting enzyme, wherein the first prodrug converting enzyme is expressed in the cytoplasm of the immune cell;(b) a second prodrug converting enzyme and/or a nucleic acid encoding the second prodrug converting enzyme, wherein the second prodrug converting enzyme is expressed on the surface of the immune cell or is secreted; and(c) a receptor that binds to a target antigen and/or nucleic acid encoding the receptor.4. The engineered immune cell of any of - claim 1 , wherein the receptor is a T cell receptor.5. The engineered immune cell of any of - claim 1 , wherein the receptor is a native receptor.6. The engineered immune cell of any of - claim 1 , wherein the receptor is a non-native receptor.7. The engineered immune cell of any of - claim 1 , wherein the receptor is a chimeric antigen receptor.8. The engineered immune cell of any of - claim 1 , wherein the second prodrug converting enzyme is fused to a transmembrane ...

NOVEL MEANS TO DECREASE THE NEGATIVE EFFECTS OF SMOKING

The present invention relates to the use of an elastase inhibitor, preferably fahsin for the treatment or prevention of emphysema, COPD or lung cancer. The elastase inhibitor is preferably administered through inhalation, preferably thorough inhalation of tobacco smoke. The invention also comprises smoking articles comprising such an elastase inhibitor. 1. Fahsin for use in the prevention of lung disease , selected from the group of emphysema , COPD and lung cancer.2. Fahsin for use in therapy or prophylaxis of lung disease , particularly emphysema , through inhalation , specifically through inhalation of smoke , more particularly tobacco smoke.3. Smoking article , such as a cigarette , including an e-cigarette , pipe tobacco , cigar or joint , for use in the therapy or prophylaxis of emphysema.4. Smoking article according to claim 3 , wherein said article comprises an elastase inhibitor claim 3 , preferably selected from the group of fahsin claim 3 , guamerin claim 3 , piguamerin claim 3 , hirustasin claim 3 , bdellastasin and mutants of guamerin claim 3 , piguamerin claim 3 , hirustasin and bdellastasin that contain a leucine residue after the 6cysteine residue.5. Smoking article according to claim 4 , wherein the elastase inhibitor is included in tobacco or cannabis claim 4 , either as a blend or as a protein expressed by said tobacco or cannabis.6. Smoking article according to claim 4 , wherein the cigarette is a filter cigarette or an e-cigarette and the elastase inhibitor is present in the filter.7. Smoking article according to claim 4 , wherein the smoking article is a cigarette or a joint and the elastase inhibitor is present in the cigarette paper.8. Smoking article according to any of - wherein the elastase inhibitor is recombinantly produced fahsin.9. Transgenic tobacco or cannabis comprising an elastase inhibitor claim 4 , preferably fahsin.10. Cigarette comprising the transgenic tobacco or cannabis according to .11. Method to prevent or reduce emphysema ...