СПОСОБЫ ПОЛУЧЕНИЯ МУЛЬТИСПЕЦИФИЧНЫХ И МУЛЬТИВАЛЕНТНЫХ АНТИТЕЛ

Изобретение относится к области биотехнологии. Предложен способ получения смеси антител, содержащей два моноспецифических антитела и одно биспецифическое антитело, где все антитела имеют общую тяжелую цепь. Выделяют два антитела с различной специфичностью, определяемой одним и тем же вариабельным доменом тяжелой цепи. Затем коэкспрессируют в клетке две различных легких цепи и одну тяжелую цепь указанных антител, что приводит к сборке моноспецифических и биспецифических антител. Одна из указанных легких цепей содержит константный домен Каппа, а другая - константный домен Лямбда. Также предложена смесь антител для выделения двух моноспецифических антител и одного биспецифического антитела, полученная указанным способом. Изобретение обеспечивает регуляцию соотношения различных антител в получаемой смеси для максимизации получения биспецифического антитела. 2 н. и 6 з.п. ф-лы, 37 ил., 5 табл., 15 пр.

АНТИТЕЛА ПРОТИВ ФАКТОРА РОСТА ЭНДОТЕЛИЯ СОСУДОВ (VEGF)

Изобретение относится к области иммунологии. Описаны антитела против VEGF, одно из которых содержит комплементарные регионы с аминокислотными последовательностями SEQ ID NO:1, 2, 3, 4, 6 и 7, другое содержит комплементарные регионы с аминокислотными последовательностями SEQ ID NO:1, 2, 3, 5, 6 и 7, раскрытыми в описании. Также описаны полинуклеотиды, кодирующие указанные антитела; экспрессионные векторы, содержащие указанные полинуклеотиды, и клетки-хозяева, предназначенные для получения антител по настоящему изобретению. Предложен способ получения антител против VEGF, включающий экспрессию вектора в клетке-хозяине и выделение антитела. Раскрыт способ получения иммуноконъюгата антитела против VEGF, включающий конъюгирование антитела с лекарством или цитотоксическим агентом. Описан способ обнаружения VEGF, включающий обнаружение комплекса VEGF-антитело против VEGF в биологическом образце. Кроме того, раскрыты композиции для лечения заболевания, ассоциированного с VEGF, одна из которых содержит ...

УЛУЧШЕННЫЕ СПОСОБЫ ОБРАЗОВАНИЯ ДИСУЛЬФИДНЫХ СВЯЗЕЙ

... 1. Способ образования дисульфидной связи, включающий: вызов или обеспечение возможности присоединения первого (поли)пептида/белка ко второму (поли)пептиду/белку, где указанное присоединение вызвано образованием дисульфидной связи между первым цистеиновым остатком, содержащимся в указанном первом (поли)пептиде/белке, и вторым цистеиновым остатком, содержащимся в указанном втором (поли)пептиде/белке, где один из указанных (поли)пептидов/белков содержит аминокислоту, имеющую рI выше 8, которая положительно влияет на реакционную способность по меньшей мере одного из указанных цистеиновых остатков. ! 2. Способ по п.1, где указанная аминокислота, имеющая рI выше 8, содержится в указанном первом (поли)пептиде/белке. ! 3. Способ по п.1, где указанная аминокислота, имеющая рI выше 8, содержится в указанном втором (поли)пептиде/белке. ! 4. Способ по п.2 или 3, где указанная аминокислота, имеющая рI выше 8, отсутствует в соответствующем аминокислотном положении в указанном (поли)пептиде/белке дикого ...

ПЕРЕКРЕСТНО-РЕАКТИВНЫЕ АНТИТЕЛА АНТИ-IL-17A/IL-17F И СПОСОБЫ ИХ ПРИМЕНЕНИЯ

... 1. Выделенное полностью человеческое моноклональное антитело, которое связывается с гомодимером IL-17A, гомодимером IL-17F и гетеродимерным комплексом IL-17A/IL-17F, при этом указанное антитело проявляет (i) аффинность связывания по меньшей мере 100 пМ или менее против гомодимера IL-17A, (ii) аффинность связывания по меньшей мере 300 пМ или менее против гомодимера IL-17F, (iii) аффинность связывания по меньшей мере 400 пМ или менее против гетеродимерного комплекса IL-17A/IL-17F, (iv) способность к нейтрализации по меньшей мере 13 нМ или менее против гомодимера IL-17A, (v) способность к нейтрализации по меньшей мере 120 нМ или менее против гомодимера IL-17F, и (vi) способность к нейтрализации по меньшей мере 31 нМ или менее против гетеродимерного комплекса IL-17A/IL-17F. ! 2. Антитело по п. 1, где антитело проявляет (i) аффинность связывания по меньшей мере 40 пМ или менее против гомодимера IL-17A, (ii) аффинность связывания по меньшей мере 10 пМ или менее против гомодимера IL-17F, и (iii ...

Иммунотерапия с применением антител, связывающих лиганд 1 белка программируемой смерти клеток (PD-L1)

СВЯЗЫВАЮЩИЕ ПОЛИПЕПТИДЫ И ИХ ПРИМЕНЕНИЯ

... 1. Полипептид, содержащий вариабельный домен тяжелой цепи иммуноглобулина, в котором ! (i) CDRH1 содержит аминокислотную последовательность G-F-X1-I-X2-X3-X4-X5-I-H (SEQ ID NO:22), где G находится в положении 26, а X1 находится в положении 28 согласно системе нумерации Кабата; где Х1 выбран из S и Y; где Х2 выбран из Y и S; где Х3 выбран из Y и S; где Х4 выбран из Y и S; и где Х5 выбран из Y и S; ! (ii) CDRH2 содержит аминокислотную последовательность: X1-I-X2-P-X3-X4-G-X5-T-X6-Y-A-D-S-V-K-G (SEQ ID NO:23), где X1 находится в положении 50 согласно системе нумерации Кабата; где Х1 выбран из Y и S; где Х2 выбран из Y и S; где Х3 выбран из Y и S; где Х4 выбран из Y и S; где Х5 выбран из Y и S; и где Х6 выбран из Y и S; и ! (iii) CDRH3 содержит аминокислотную последовательность: X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14-X15-X16-D-Y (SEQ ID NO:31), где X1 находится в положении 95 согласно системе нумерации Кабата, и где Х1 выбран из R, Y и M; Х2 выбран из Y и R; Х3 выбран из Y, S, R, P ...

MULTIVALENTE UND MULTISPEZIFISCHE BINDUNGSPROTEINE, DEREN HERSTELLUNG UND VERWENDUNG

Anti-VEGF Antik¦rper.

ANTI-VEFG ANTIBODIES

Screening methods

Retargeting

ANTIBODIES

BINDING SUBSTANCE

Primers

Anti-VEGF antibodies.

BACTERIOPHAGE PARTICLES, THE DABS PRESENTATION

ARTIFICIAL ANTI-BODY LIBRARY WITH SUPER REPERTOIRE

PERFECTLY HUMAN ANTIBODIES THE EGFR BINDING

SELFARRANGING MOLECULES

ARRANGED LIBRARIES ARE GENETICALLY PACKED

ANTIGEN-BINDING DOMAIN FROM FISH

MULTIVALENTE IMMUNGLOBULINE

The present invention provides a multivalent immunoglobulin or part thereof binding specifically to at least two cell surface molecules of a single cell with at least one modification in at least one structural loop region of said immunoglobulin determining binding to an epitope of said cell surface molecules wherein the unmodified immunoglobulin does not significantly bind to said epitope, its use and methods for producing it.

FAB FRAGMENT LIBRARIES AND PROCEDURES FOR THEIR USE

CONTAINER WITH ANTI- VEGF ANTIBODIES

PEGYLIERTE SINGLE DOMAIN ANTIKÖRPER ONE (DAB)

PROCEDURE FOR THE SIFTING OF PHAGE DISPLAY LIBRARIES WITH DIFFERENT LIGANDS

PROCEDURE FOR THE PRODUCTION OF FUNCTIONAL, EINKETTIGER FV OF ANTIKOERPER FRAGMENTS ON BACTERIOPHAGE SURFACES

SCREENING PROCEDURE FOR GENE BANKS (RECOMBINANT LIBRARIES)

RATIONALLY SKETCHED ANTIBODIES

PRODUCTION OF ANTIBODIES ON PHAGE SURFACES ON THE BASIS OF ANTI-BODY SEGMENT LIBRARIES.

METHOD FOR THE PRODUCTION OF FILAMENT EYES, ANTI-BODY-PRESENTING BACTERIOPHAGE PARTICLES BY USE OF PHAGEMIDEN AND THE DADUCH MANUFACTURE BACTERIOPHAGE PARTICLE

MULTIVALENTE AND MULTI-SPECIFIC CONNECTION PROTEINS, THEIR PRODUCTION AND USE

PURPOSEFUL ANTI-BODY PROCEDURE

Binding agents

The invention relates to antibody molecules and antigen-binding portions thereof which bind specifically to glucocorticoid-induced TNF receptor (GITR). In particular aspects of the invention, the antibody molecules specifically bind to human GITR and cynomolgus monkey GITR. The anti-GITR antibody molecules of the invention have been developed and optimized using CDR sequences derived from a murine anti-GITR antibody 6C8. Medical uses of the antibody molecules are disclosed.

Ion concentration-dependent binding molecule library

ION CONCENTRATION-DEPENDENT BINDING MOLECULE LIBRARY Disclosed is a library primarily comprising a plurality of antigen-binding molecules having different sequences and having at least one amino acid residual contained in the antigen-binding domain, said amino acid residual varying the binding activity of the antigen binding molecules with respect to the antigens, according to the ion concentration conditions. Also disclosed are: a composition containing a plurality of polynucleotide molecules that code antigen-binding molecules; a composition containing a plurality of vectors that contain the aforementioned polynucleotide molecules; a method for selecting the aforementioned antigen binding molecule; a method for isolating the aforementioned polynucleotide molecules; a method for preparing the aforementioned antigen-binding molecules; and a pharmaceutical composition containing the aforementioned antigen-binding molecules.

Dynamic human antibody light chain libraries

Provided herein are libraries containing polynucleotides, where one of the polynucleotides encodes an antibody light chain with specific hypervariable regions HVR-L1, HVR-L2, and HVR-L3. Further provided herein are libraries containing polynucleotides encoding a plurality of unique antibodies, wherein each antibody comprises a heavy chain variable region and a light chain variable region. Also provided are antibodies, polypeptide libraries, vector libraries, cells, non-human animals, antibody light chains, methods of making an antibody library, kits, and methods of generating a bispecific antibody related thereto.

Mixed binding domains

A binding domain or a multimer or a variant thereof which comprises a variable region encoded by a nucleic acid based on, derived or obtained from an animal phylogenetically distal from a human, which variable region is paired with a human variable region.

Antibody and method for producing same

Disclosed is an antibody in which the 80th amino acid residue in a variable region based on the Kabat method and the 171th amino acid residue in a constant region based on the Kabat method are substituted with cysteine in an antibody in which the 80th amino acid residue in the variable region and the 171th amino acid residue in the constant region are not cysteine.

Anti-TAT226 antibodies and immunoconjugates

Methods and compositions for targeting polyubiquitin

Engineered antibody constant domain molecules

Polypeptides, antibody variable domains and antagonists

Polypeptides, antibody variable domains and antagonists

High affinity anti-human IgE antibodies

Expression of biologically active polypeptides in duckweed

Solution phase biopanning method using engineered decoy proteins

Methods for the generation of multispecific and multivalent antibodies

The invention provides novel bispecific monoclonal antibodies carrying a different specificity for each binding site of the immunoglobulin molecule and methods for producing novel bispecific monoclonal antibodies carrying a different specificity for each binding site of the immunoglobulin molecule. The antibodies are composed of a single heavy chain and two different light chains, one containing a Kappa constant domain and the other of a Lambda constant domain. The invention provides methods for the isolation of antibodies of different specificities but sharing a common heavy chain. The invention also provides methods for the controlled co-expression of two light chains and a single heavy chain leading to the assembly of monospecific and bispecific antibodies. The invention provides a mean of producing a fully human bispecific and bivalent antibody that is unaltered in sequence and does not involve the use of linkers or other non-human sequences, as well as antibody mixtures of two monospecific ...

Compositions and methods relating to anti-IGF-1 receptor antibodies

The present invention provides compositions and methods relating to or derived from anti-IGF-IR antibodies. In particular embodiments, the invention provides fully human, humanized, or chimeric anti-IGF-lR antibodies that bind human IGF-lR, IGF-IR-binding fragments and derivatives of such antibodies, and IGF-IR 5 binding polypeptides comprising such fragments. Other embodiments provide nucleic acids encoding such antibodies, antibody fragments and derivatives and polypeptides, cells comprising such polynucleotides, methods of making such antibodies, antibody fragments and derivatives and polypeptides, and methods of using such antibodies, antibody fragments and derivatives and polypeptides, including methods of treating or diagnosing subjects having IGF- I R-related disorders or conditions.

Synthetic Polypeptide Libraries And Methods For Generating Naturally Diversified Polypeptide Variants

The invention provides compositions and methods for generating libraries of DNA sequences encoding homologous polypeptides, and uses of the libraries to identify naturally diversified polypeptide variants. The invention also provides compositions and methods for generating collections of synthetic antibody fragments in which one or several complementary determining regions (CDR) are replaced by a collection of the corresponding CDR captured from a natural source. The invention further provides compositions and methods for diversifying a portion of a polypeptide by inserting a diversified sequence of synthetic or natural origin without the need for modification of the original polypeptide coding sequence.

Novel methods of constructing libraries comprising displayed and/or expressed members of a diverse family of peptides, polypeptides or proteins and the novel libraries

Comprehensive monoclonal antibody generation

The present invention relates to methods for efficiently generating recombinant monoclonal antibodies derived from B cells of a non-human host which has been immunochallenged with one or more target antigens. The methods comprise the steps of identifying and isolating B cell that bind to the antigen by FACS, and recombining and enriching for thousands of cells to create a B cell library. Related products and methods, such as methods of producing expression libraries, are also disclosed.

Antibodies against immunocomplexes comprising cyanobacterial cyclic peptide hepatotoxins

The present invention relates to means and methods for detecting cyanobacterial cyclic peptide hepatotoxins (CCPH) in aqueous samples, More specifically, the invention provides recombinant anti-immunocomplex (anti-IC) antibodies which bind to immunocomplexes formed between one or more CCPH variants and an anti-CCPH primary antibody, and immunoassays, preferably non-competitive immunoassays, employing the same.

Multi-specific monoclonal antibodies

MULTI-SPECIFIC MONOCLONAL ANTIBODIES Abstract The present invention is relevant to the generation of multi-specific antibodies, antibodies that are distinguished by their ability to bind multiple antigens with specificity and with affinity. In particular, the present invention is related to bi specific antibodies.

Compositions and methods for growth factor modulation

Provided herein are proteins, antibodies, assays and methods useful for modulating growth factor levels and/or activities. In some embodiments, such growth factors are members of the TGF-p superfamily of proteins.

PRODUCTION OF ANTI-SELF ANTIBODIES FROM ANTIBODY SEGMENT REPERTOIRES AND DISPLAYED ON PHAGE

EXPRESSION OF BIOLOGICALLY ACTIVE POLYPEPTIDES IN DUCKWEED

ANTIBODY COMPOSITIONS AND METHODS

Provided are concentrated preparations comprising single immunoglobulin variable domain polypeptides that bind target antigen with high affinity and are soluble at high concentration, without aggregation or precipitation, providing, for example, for increased storage stability and the ability to administer higher therapeutic doses.

ANTIBODIES AGAINST CXCR4 AND METHODS OF USE THEREOF

... ²²²The invention provides human monoclonal antibodies, scFv antibodies, scFv-Fc ²fusions, a dAb (domain antibodies), Fab, Fab' and F(ab')2 fragments, single ²chain antibodies and/or minibodies that specifically bind to CXCR4. Also ²provided are methods of treating and/or preventing a CXCR4 disease or disorder ²such as cancer and X4-tropic HIV-I infection as well as uses of such ²antibodies and antibody fragments in the manufacture of a medicament for the ²treatment or prevention of a CXCR4 disorder. The invention also provides ²methods of preventing diseases or disorders associated with CXCR4 function or ²expression. The invention further provides for the use of the antibodies (or ²fragments thereof) of the invention in the manufacture of a medicament for the ²prevention of diseases or disorders associated with CXCR4 expression. Also ²provided are methods of treating or preventing cancer metastasis in a patient ²suffering from a cancer involving tumor cells that express CXCR4. The ²invention ...

INTRABODIES WITH DEFINED FRAMEWORK THAT IS STABLE IN A REDUCING ENVIRONMENT AND APPLICATIONS THEREOF

... ²²²A method for the isolation of CDRs in a defined framework that is stable and ²soluble in reducing environment is described as well as thus obtainable scFv. ²Starting from such scFv with defined framework a scFv library can be generated ²wherein the framework is conserved while at least one complementary ²determining region (CDR) is randomized. Such library, e.g. in yeast cells, is ²suitable for screening for antibody/CDR-interactions or for screening for ²antibodies.² ...

METHODS FOR SELECTING PROTEASE RESISTANT POLYPEPTIDES

The invention relates to a method for selecting, isolating and/or recover ing a peptide or polypeptide from a library or a repertoire of peptides and polypeptides ( e.g., a display system) that is resistant to degradation by a protease such as a protease found in the GI tract or pulmonary tissue of a human. Generally, the method comprises providing a library or repertoire of peptides or polypeptides, combining the library or repertoire with a proteas e under conditions suit able for protease activity, and selecting, isolating and/or recovering a peptide or polypeptide that is resistant to degradation by the protease and has a desired biological activity. The selected peptide s and polypeptides have utility as therapeutics, eg for treating disease or conditions of GI tract or pulmonary tissue in humans.

IMPROVED IMMUNOGLOBULIN LIBRARIES

Methods and compositions for the screening and isolation of ligand-binding polypeptides, such as antibodies. In some aspects, methods of the invention enable the isolation of intact soluble antibodies comprising a constant domain. Screening methods that employ genetic packages such as bacteria and bacteriophages enable high through-put identification of ligand binding molecules.

IMPROVED METHODS FOR THE FORMATION OF DISULPHIDE BONDS

The present invention relates to methods for the formation of inter-molecular disulphide bonds, including (poly)peptides/proteins, nucleic acids, vectors, host cells and bacteriophages used in these methods. Furthermore the invention relates to the use of this method for the improved display of (poly)peptides/proteins on the surface of bacteriophage particles.

ANTIGEN BINDING DOMAINS

A process for the production of an antigen specific antigen binding domain using a transformed host containing an expressible DNA sequence encoding the antigen specific antigen binding domain, wherein the antigen specific antigen binding domain is derived from a variable region of the immunoglobulin isotype NAR found in fish.

METHODS AND COMPOSITIONS FOR DISCOVERY OF TARGET-SPECIFIC ANTIBODIES USING ANTIBODY REPERTOIRE ARRAY (ARA)

The invention provides antibody arrays specific for target antigens. Methods for discovery and compositions comprising native human antibodies, arrays comprising such antibodies, immortalized B cells expressing such antibodies and non-immortalized B cell libraries comprising B cells expressing such antibodies are provided. The invention provides a method for screening monoclonal antibodies for functional effects on cell surface molecules such as receptors using antibody repertoire arrays specific for target cell surface molecules. Functional antibodies directed to a target and therapeutics derived from such antibodies are also provided. High throughput and parallel screening for potentially therapeutic antibodies are provided. Antibodies directed to functional epitope clusters corresponding to a target and vaccines and therapeutics derived from such antibodies are also provided.

ANTIBODY CDR POLYPEPTIDE SEQUENCES WITH RESTRICTED DIVERSITY

The invention provides variant CDRs comprising highly restricted amino acid sequence diversity. These polypeptides provide a flexible and simple source of sequence diversity that can be used as a source for identifying novel antigen binding polypeptides. The invention also provides these polypeptides as fusion polypeptides to heterologous polypeptides such as at least a portion of phage or viral coat proteins, tags and linkers. Libraries comprising a plurality of these polypeptides are also provided. In addition, methods of and compositions for generating and using these polypeptides and libraries are provided.

RATIONALLY DESIGNED ANTIBODIES

Antibodies or fragments thereof having at least two CDR regions replaced or fused with biologically active peptides are described. Compositions containing such antibodies or fragments thereof are useful in therapeutic and diagnostic modalities.

POLYPEPTIDE INCLUDING ANTIGEN-BINDING DOMAIN AND CARRYING SECTION

The present invention relates to: a polypeptide that includes an antigen-binding domain and a carrying section having an inhibitory domain for inhibiting the antigen-binding activity of the antigen-binding domain, said polypeptide having a half-life longer than that of the antigen-binding domain when existing alone; a production method and a screening method for said polypeptide; a pharmaceutical composition containing said polypeptide; a production method and a screening method for a single-domain antibody, the antigen-binding activity of which is inhibited by being associated with a specific VL/VH/VHH; and a library of fused polypeptides including a single-domain antibody, the antigen-binding activity of which is inhibited by being associated with a specific VL/VH/VHH.

AMYLOID-.BETA.(1-42) OLIGOMERS, DERIVATIVES THEREOF, ANTIBODIES FOR THE SAME, METHOD FOR PRODUCTION AND USE THEREOF

... ²²The invention relates to neuromodulatory oligomers of the amyloid-.beta.(1-42) ²protein, a particular ²production method, by means of which the oligomer can be obtained in a ²reproducible manner at ²high yield, the use of the oligomers as diagnostic and therapeutics agents, ²for the generation of ²oligomer-specific antibodies and for the discovery of substances which can ²interact with the ²oligomers and in the formation thereof. Corresponding methods for the ²production of the ²antibodies and for discovery of the substances are also disclosed as are the ²antibodies themselves ²and the use of the antibodies or substances as diagnostic and therapeutic ²agents. The invention ²further relates to derivatives of the oligomers and oligomers based on ²abbreviated forms of the ²amyloid-.beta.(1-42) proteins, the production and use thereof.² ...

CONDITIONALLY ACTIVE POLYPEPTIDES AND METHODS OF GENERATING THEM

A method of preparing a conditionally active polypeptide from a parent polypeptide, comprising steps of evolving a DNA encoding the parent polypeptide by increasing a net charge of the parent polypeptide using one or more techniques selected from increasing a total number of codons of charged amino acid residues in the DNA and decreasing a total number of codons of uncharged amino acid residues in the DNA to create mutant DNAs; expressing the mutant DNAs to obtain mutant polypeptides; and selecting the conditionally active polypeptide from the mutant polypeptides which exhibits a decrease in activity in a first assay at a first value of a condition compared to the same activity in a second assay at a second value of the same condition. The conditionally active polypeptide, pharmaceutical compositions containing same, nanoparticle and drug conjugates thereof and uses thereof are also provided.

DYNAMIC HUMAN HEAVY CHAIN ANTIBODY LIBRARIES

Provided herein are libraries containing polynucleotides, where one of the polynucleotides encodes an antibody heavy chain with specific hypervariable regions HVR-H1 and HVR-H2. Further provided herein are libraries containing polynucleotides encoding a plurality of unique antibodies, wherein each antibody comprises a heavy chain variable region and a light chain variable region. Also provided are antibodies, polypeptide libraries, vector libraries, cells, non-human animals, antibody heavy chains, methods of making an antibody library, kits, and methods of generating a bispecific antibody related thereto.

MULTISPECIFIC PROTEIN DRUG AND LIBRARY THEREOF, PREPARING METHOD THEREFOR AND APPLICATION THEREOF

Provided are a multispecific protein drug and a library thereof, a preparing method therefor and an application thereof. Specifically, a protein drug library is provided and comprises C different protein drug monomers, wherein the protein drug monomer comprises a protein drug component part and a nucleic acid component part connected with the protein drug component part, and the nucleic acid component part of one protein drug monomer establishes a double-stranded paired structure with a nucleic acid component part of at least one different protein drug monomer by means of complementation, thereby constituting a protein drug polymer, wherein C is a positive integer greater than or equal to 2.

METHOD FOR THE CONSTRUCTION OF RANDOMIZED GENE SEQUENCE LIBRARIES IN CELLS

An in vivo method for the construction of randomized gene libraries and/or domain replacement in gene libraries by homologous recombination using a Kluyveromyces lactis killer toxin, in particular the (.gamma.-subunit of the K. lactis killer toxin, as negative selection marker is described. The use of the (.gamma.-subunit of K. lactis as negative selectable marker increases the percentage of randomized clones.

METHOD FOR THE CONSTRUCTION OF RANDOMIZED GENE SEQUENCE LIBRARIES IN CELLS

An in vivo method for the construction of randomized gene libraries and/or domain replacement in gene libraries by homologous recombination using a Kluyveromyces lactis killer toxin, in particular the (.gamma.-subunit of the K. lactis killer toxin, as negative selection marker is described. The use of the (.gamma.-subunit of K. lactis as negative selectable marker increases the percentage of randomized clones.

ANTIBODY POLYPEPTIDE LIBRARY SCREENING AND SELECTED ANTIBODY POLYPEPTIDES

METHOD FOR PREPARING IMMUNOGLOBULIN LIBRARIES

The invention relates to methods of construction of human immunoglobulin libraries, by isolating B memory cells (IgM memory, class-switched or plasma blast cells) . The libraries are generated on the surface of replicable genetic packages, such as phages. It also relates to methods of screening said libraries and the immunoglobulins identified in said screenings. Additionally, the invention relates to human binding molecules that bind to H5N1 virus, in particular antibodies and immunoconjugates. Pharmaceutical compositions comprising said binding molecules are also disclosed.

ANTIBODY ULTRAHUMANIZATION BY PREDICTED MATURE CDR BLASTING AND COHORT LIBRARY GENERATION AND SCREENING

ANTI-VEGF ANTIBODIES

... ²Anti-VEGF antibodies and variants thereof, including those having high ²affinity for ²binding to VEGF, are disclosed. Also provided are methods of using phase ²display ²technology with naïve libraries to generate and select the anti-VEGF ²antibodies with desired ²binding and other biological activities. Further contemplated are uses of the ²antibodies in ²research, diagnostic and therapeutic applications.²² ...

CONDITIONALLY ACTIVE BIOLOGICAL PROTEINS

Methods of generating conditionally active biologic proteins, in particular therapeutic or diagnostic proteins, which are more active at an aberrant condition than at a normal physiological condition. The methods include discovery methods using libraries of proteins and assays employing physiological concentrations of components of bodily fluids. The conditionally active biologic proteins may be further evolved, conjugated to other molecules, masked, reduced in activity by attaching a cleavable moiety. Criteria for selecting starting proteins for the discovery methods, as well as formats of the proteins are also disclosed.

METHODS FOR NON-COVALENT FC-DOMAIN-CONTAINING PROTEIN DISPLAY ON THE SURFACE OF CELLS AND METHODS OF SCREENING THEREOF

The present invention provides methods for the non-covalent surface display of proteins of interest (POI), in particular for Fc-domain containing proteins such as antibodies. The inventive method may be used to screen and select proteins of interest of a desired phenotype. The present invention further discloses polynucleotides and proteins and methods of producing the same, which may be used in carrying out the inventive method.

HIGH THROUGHPUT SCREENING METHOD AND USE THEREOF TO IDENTIFY A PRODUCTION PLATFORM FOR A MULTIFUNCTIONAL BINDING PROTEIN

Methods of identifying and expressing an antibody variant are disclosed wherein the method comprises identifying a binding region in an antibody, fusing the binding region to a plurality of scaffolds of antibody constant regions to obtain antibody fragment variants, expressing the antibody fragment variants in organisms to form constructs and expressing the constructs carried by the organisms to form induced cultures, wherein the organisms are expressed in HTP mode.

METHOD OF ISOLATION OF SOLUBLE POLYPEPTIDES

Polypeptides with desirable biophysical properties such as solubility, stability, high expression, monomericity, binding specificity or non-aggregation, including monomeric human VHs and VLs, are identified using a high throughput method for screening polypeptides, comprising the steps of obtaining a phage display library, allowing infection of a bacterial lawn by the library phage, and identifying phage which form larger than average plaques on the bacterial lawn. Sequences of monomeric human VHs and VLs are identified, which may be useful for immunotherapy or as diagnostic agents. Multimer complexes of human VHs and VLs are also identified. The VHs and VLs identified may be used to create further libraries for identifying additional polypeptides. Further, the VHs and VLs may be subjected to DNA shuffling to select for improved biophysical properties.

METHOD FOR PRODUCING SECRETABLE ANTIBODIES BY EXPRESSION IN SACCHAROMYCES CEREVISIAE

The invention relates to a method for the production and non-covalent surface display of antibodies and derived fragments as well as molecule libraries based thereon on the surface of S. cerevisiae cells. The non-covalent manner of the surface display enables specific variants to be selected by means of high-throughput screening and the selected binding molecule to be subsequently controllably secreted into the culture supernatant for biochemical characterization.

MULTIVALENT BINDING PROTEIN COMPOSITIONS

Provided are multivalent binding proteins that specifically bind to one or more desired target antigens. The disclosure also provides nucleic acid, vectors and host cells encoding multivalent binding proteins, methods of producing and methods of using and the multivalent binding proteins.

PRODUCTION OF CHIMERIC ANTIBODIES - A COMBINATORIAL APPROACH

... 2119930 9306213 PCTABS00021 Methods are disclosed which may be used for the production of antibodies, or antibody fragments, which have the same binding specificity as a parent antibody but which have increased human characteristics. Humanised antibodies may be obtained by chain shuffling, perhaps using phage display technology. In one embodiment, a polypeptide comprising a heavy or light chain variable domain of a non-human antibody specific for an antigen of interest is combined with a repertoire of human complementary (light or heavy) chain variable domains. Hybrid pairings which are specific for the antigen of interest are selected. Human chains from the selected pairings may then be combined with a repertoire of human complementary variable domains (heavy or light) and humanised antibody polypeptide dimers can then be selected for binding specificity for antigen. The methods may be combined with CDR-imprinting. In another embodiment, component part of an antigen-binding site of a non-human ...

PROTEIN/(POLY)PEPTIDE LIBRARIES

The present invention relates to synthetic DNA sequences which encode one or more collections of homologous proteins/(poly)peptides, and methods for generating and applying libraries of these DNA sequences. In particular, the invention relates to the preparation of a library of human-derived antibody genes by the use of synthetic consensus sequences which cover the structural repertoire of antibodies encoded in the human genome. Furthermore, the invention relates to the use of a single consensus antibody gene as a universal framework for highly diverse antibody libraries.

SINGLE BLAST-FURNACE DOMAIN PROTEIN, SERUM ALBUMIN BINDING

УЛУЧШЕННЫЕ СВЯЗЫВАЮЩИЕ МОЛЕКУЛЫ НА ОСНОВЕ ФИБРОНЕКТИНА И ИХ ПРИМЕНЕНИЕ

В заявке описаны основанные на фибронектине III типа (Fn3) связывающие молекулы, которые связываются со специфическим антигеном-мишенью. В заявке также описаны биспецифические основанные на Fn3 связывающие молекулы, которые связываются с двумя или несколькими мишенями одновременно. Основанные на Fn3 связывающие молекулы по настоящему изобретению могут также быть связаны вместе для формирования полиспецифических основанных на Fn3 связывающих молекул и/или могут быть конъюгированы с частью молекулы, не являющейся Fn3, например с сывороточным альбумином человека (САЧ), для улучшения периода полураспада и стабильности. В заявке также описаны способы получения, скрининга и применения основанных на Fn3 связывающих молекул в различных терапевтических и диагностических применениях.

RECOMBINANT ANTIBODY AGAINST THE RH FACTOR D THE METHODS OF ITS OBTAINING

POLYPEPTIDE, THE VARIABLE DOMAINS OF ANTIBODIES AND THE ANTAGONISTS

ПОЛИПЕПТИДЫ, ВАРИАБЕЛЬНЫЕ ДОМЕНЫ АНТИТЕЛ И АНТАГОНИСТЫ

Изобретение относится к полипептидам и единичным вариабельным доменам (dAb) антител против TNFR1, устойчивым к деградации протеазой, а также к содержащим их антагонистам. Полипептиды, dAb и антагонисты полезны в качестве терапевтических и/или профилактических средств, которые вероятно сталкиваются с протеазами при введении пациенту, например для внутрилегочного введения, перорального введения, доставки в легкое и доставки в ЖК тракт пациента, а также для лечения воспалительного заболевания, такого как артрит или COPD (хроническое обструктивное заболевание легких).

ПОЛИПЕПТИДЫ, ВАРИАБЕЛЬНЫЕ ДОМЕНЫ АНТИТЕЛ И АНТАГОНИСТЫ

Настоящее изобретение относится к единичным вариабельным доменам иммуноглобулинов (dAb), например dAb, которые являются протеазоустойчивыми, и также к препаратам и композициям, содержащим такие dAb, для доставки в глаз и к их применениям для лечения глазных заболеваний и состояний.

ПОЛИПЕПТИДЫ, ВАРИАБЕЛЬНЫЕ ДОМЕНЫ АНТИТЕЛА И АНТАГОНИСТЫ

Изобретение относится к полипептидам и единичным вариабельным доменам (dAb) антитела против IL-1R1, которые устойчивы к деградации протеазой, а также содержащим их антагонистам. Указанные полипептиды, dAb и антагонисты полезны в качестве терапевтических и/или профилактических средств, которые, вероятно, сталкиваются с протеазами при введении пациенту, например для внутрилегочного введения, перорального введения, доставки в легкое и доставки в ЖК (желудочно-кишечный) тракт пациента, а также для лечения воспалительного заболевания, такого как артрит или COPD (хроническое обструктивное заболевание легких).

Construction method and application of chimeric antigen receptor (CAR) based on BMCA nano-antibody sequence

Help to choose the specific immunoglobulin gene recombinant vaccinia virus infection of the cells of the fusion protein

POLYPEPTIDE VARIANTS WITH ALTERED EFFECTOR FUNCTION

The invention provides polypeptides having IgG Fc regions with amino acid modifications that result in the polypeptides exhibiting altered Fc effector functions. © KIPO & WIPO 2008 ...

ANTIGEN-BINDING MOLECULES, THE ANTIGEN-BINDING ACTIVITY OF WHICH VARIES ACCORDING TO THE CONCENTRATION OF COMPOUNDS, AND LIBRARIES OF SAID MOLECULES

ENGINEERING OF IMMUNOGLOBULIN DOMAINS

Constructs and libraries comprising antibody surrogate light chain sequences

The invention concerns constructs and libraries comprising antibody surrogate light chain sequences. In particular, the invention concerns constructs comprising VpreB sequences, optionally partnered with another polypeptide, such as, for example, antibody heavy chain variable domain sequences, and libraries containing the same.

CH2 Domain Template Molecules Derived From Rational Grafting Of Donor Loops Onto CH2 Scaffolds

Novel CH2 domain template molecules wherein donor loops from a database of domains are transferred to a CH2 domain scaffold. At least one or up to three loops from a donor are transferred to the CH2 domain. The donor loops may be chosen based on length, e.g., the donor loop may have a length that is similar to that of a structural loop in the CH2 domain scaffold.

Compositions and methods relating to anti-igf-1 receptor antibodies

The present invention provides compositions and methods relating to or derived from anti-IGF-1R antibodies. In particular embodiments, the invention provides fully human, humanized, or chimeric anti-IGF-1R antibodies that bind human IGF-1R, IGF-1R-binding fragments and derivatives of such antibodies, and IGF-1R-binding polypeptides comprising such fragments. Other embodiments provide nucleic acids encoding such antibodies, antibody fragments and derivatives and polypeptides, cells comprising such polynucleotides, methods of making such antibodies, antibody fragments and derivatives and polypeptides, and methods of using such antibodies, antibody fragments and derivatives and polypeptides, including methods of treating or diagnosing subjects having IGF-1R-related disorders or conditions.

Donor specific antibody libraries

The present invention concerns donor-specific antibody libraries derived from a patient donor who has suffered from, or is suffering from one or more diseases discussed herein. The present invention also concerns the method of making and using the donor-specific antibodies. The present invention further concerns the neutralizing antibodies obtained from the donor-specific antibody libraries and the methods of using these antibodies for the prevention/treatment of human disease.

Focused libraries of genetic packages

Focused libraries of vectors or genetic packages that display, display and express, or comprise a member of a diverse family of antibody peptides, polypeptides or proteins and collectively display, display and express, or comprise at least a portion of the focused diversity of the family. The libraries have length and sequence diversities that mimic that found in native human antibodies.

Anti-Rhesus D Recombinant Polyclonal Antibody and Methods of Manufacture

The invention relates to a method for manufacturing an anti-RhD recombinant polyclonal antibody composition (anti-RhD rpAb). The method comprises obtaining a collection of cells transfected with a library of anti-RhD antibody expression vectors, wherein each cell in the collection is capable of expressing from a VH and VL comprising nucleic acid segment, one member of the library, which encodes a distinct member of anti-RhD recombinant polyclonal antibody composition and is located at the same site in the genome of individual cells in said collection. The cells are cultured under suitable conditions for expression of the recombinant polyclonal antibody, which is obtained from the cells or culture supernatant. Nucleic acid segments encoding the anti-RhD rpAb are introduced into the cells by transfection with a library of vectors for site-specific integration. The method is suitable for manufacturing anti-RhD rpAb, thereby making available a superior replacement of plasma-derived prophylactic and therapeutic immunoglobulin products.

METHOD FOR ISOLATION OF SOLUBLE POLYPEPTIDES

Polypeptides with desirable biophysical properties such as solubility, stability, high expression, monomericity, binding specificity or non-aggregation, including monomeric human Vs and Vs, are identified using a high throughput method for screening polypeptides, comprising the steps of obtaining a phage display library, allowing infection of a bacterial lawn by the library phage, and identifying phage which form larger than average plaques on the bacterial lawn. Sequences of monomeric human Vs and Vs are identified, which may be useful for immunotherapy or as diagnostic agents. Multimer complexes of human Vs and Vs are also identified. The Vs and Vs identified may be used to create further libraries for identifying additional polypeptides. Further, the Vs and Vs may be subjected to DNA shuffling to select for improved biophysical properties. 123.-. (canceled)24. A polypeptide having an amino acid sequence selected from the group consisting of: SEQ ID NO:8-54.25. (canceled)26. A nucleic acid sequence that encodes a polypeptide as claimed in .2749-. (canceled)50. A multimer comprising at least two Vantibody fragments selected from SEQ ID NOs:8-22 claim 24 , or at least two Vantibody fragments selected from SEQ ID NOs:23-54.51. (canceled)52. A multimer comprising at least one Vantibody fragment selected from SEQ ID NOs:8-22 claim 24 , and at least one Vantibody fragment selected from SEQ ID NOs:23-54.5396.-. (canceled)97. A pharmaceutical composition comprising the polypeptide sequence of and a pharmaceutically suitable agent.9899.-. (canceled) This application is a division of U.S. patent application Ser. No. 11/887,113 issued as U.S. Pat. No. 8,293,233, which claims the benefit of PCT Application No. PCT/CA20061000451, which claims priority to U.S. Provisional Patent Application No. 60/664,954.The sequence listing is provided herewith in electronic form under the file name 20121206_sequence_listing.txt, created on Dec. 6, 2012, with a size of 56,279 bytes, and is ...

Anti-vegf antibodies

Anti-VEGF antibodies and variants thereof, including those having high affinity for binding to VEGF, are disclosed. Also provided are methods of using phage display technology with naïve libraries to generate and select the anti-VEGF antibodies with desired binding and other biological activities. Further contemplated are uses of the antibodies in research, diagnostic and therapeutic applications.

Fusion Proteins to Facilitate Selection of Cells Infected with Specific Immunoglobulin Gene Recombinant Vaccinia Virus

The present invention relates to a high efficiency method of expressing immunoglobulin molecules in eukaryotic cells. The invention is further drawn to a method of producing immunoglobulin heavy and light chain libraries, particularly using the trimolecular recombination method, for expression in eukaryotic cells. The invention further provides methods of selecting and screening for antigen-specific immunoglobulin molecules, and antigen-specific fragments thereof. The invention also provides kits for producing, screening and selecting antigen-specific immunoglobulin molecules. Finally, the invention provides immunoglobulin molecules, and antigen-specific fragments thereof, produced by the methods provided herein. 1. A fusion protein comprising (a) a first polypeptide segment comprising a heavy chain constant region domain and (b) a second polypeptide segment comprising the transmembrane domain of a vaccinia extracellular enveloped virus (EEV)-specific membrane protein.2. The fusion protein of claim 1 , further comprising a third polypeptide segment comprising an immunoglobulin heavy chain variable region or fragment thereof.3. The fusion protein of claim 2 , further comprising a signal peptide for facilitating expression of the fusion polypeptides on the surface of EEV.4. The fusion protein of claim 1 , wherein the vaccinia EEV-specific membrane protein is A56R.5. The fusion protein of claim 1 , wherein the second polypeptide segment further comprises the extracellular domain of the EEV-specific membrane protein claim 1 , or a portion thereof.6. The fusion protein of claim 1 , wherein the second polypeptide segment further comprises the intracellular domain of the EEV-specific membrane protein claim 1 , or a portion thereof.7. The fusion protein of claim 1 , wherein the constant region comprises a CH1 domain claim 1 , or a portion thereof.8. The fusion protein of claim 1 , wherein the constant region comprises a IgG-gamma heavy chain.9. The fusion protein of claim 1 ...

ENGINEERING OF IMMUNOGLOBULIN DOMAINS

The present invention provides a single domain antibody (sdAb) scaffold comprising one or more than one non-canonical disulfide bond in the framework region (FR). The one or more than one non-canonical disulfide bond may be formed between cysteines introduced by mutations in FR2 and FR3. In the case where the sdAb scaffold is a V, the Cys may be introduced at any one of positions (47-49) and any one of positions (67-71), based on Kabat numbering; in one example, the Cys may be introduced at positions (49) and (69), based on Kabat numbering. In the case where the sdAb scaffold is a V, the Cys residues may be introduced at any one of positions 46-49 and any one of positions (62-66), based on Kabat numbering; in one example, the Cys residues may be introduced at positions (48 and 64), based on Kabat numbering. 1. A composition comprising an immunoglobulin scaffold comprising one or more than one non-canonical disulfide bond in the framework region (FR).2. (canceled)3. The composition of claim 1 , wherein the scaffold is part of a larger antibody protein or fragment selected from the group consisting of a single domain antibody (sdAb) claim 1 , scFv claim 1 , Fab claim 1 , F(ab) claim 1 , or mature immunoglobulin.4. The composition of claim 3 , wherein the mature immunoglobulin is selected from the group consisting of an IgG1 claim 3 , IgG2 claim 3 , IgG3 claim 3 , IgG4 claim 3 , IgE claim 3 , and/or IgM.5. The composition of claim 1 , wherein the immunoglobulin scaffold is a V.6. The composition of claim 5 , wherein the Vis of the V3 family.7. The composition of claim 1 , wherein the immunoglobulin scaffold is a V.8. The composition of claim 7 , wherein the Vis of the kappa or lambda family.9. The composition of claim 1 , wherein the one or more than one non-canonical disulfide bond is formed between cysteine residues introduced by mutations into FR2 and FR3.10. The composition of claim 5 , wherein the cysteine residues are introduced at positions 49 and 69 claim 5 , ...

Human TIMP-1 Antibodies

Human antibodies that bind to TIMP-1 can be used as reagents to diagnose and treat disorders in which TIMP-1 is elevated, such as liver fibrosis, alcoholic liver disease, cardiac fibrosis, acute coronary syndrome, lupus nephritis, glomerulosclerotic renal disease, benign prostate hypertrophy, colon cancer, lung cancer, and idiopathic pulmonary fibrosis.

Cytoplasmic Expression of Fab Proteins

The present invention relates generally to antigen binding polypeptides, such as Fab fragments and derivatives thereof, that demonstrate high stability and solubility. The present invention also relates to polynucleotides encoding such polypeptides, to libraries of such polypeptides or polynucleotides, and to methods of using such polypeptides in research, diagnostic and therapeutic applications. For example, the polypeptides can be used in screening methods to identify a polypeptide that binds to a particular target molecule. 147.-. (canceled)48. A polynucleotide library comprising a plurality of different polynucleotides , wherein each polynucleotide encodes a Fab fragment or derivative thereof comprising:a. an antibody heavy chain variable region (VH) comprising a scaffold region which is at least 90% identical to the scaffold region of IGHV3-23 as set out in SEQ ID NO: 3; andb. an antibody light chain variable region (VL) comprising a scaffold region which is at least 90% identical to the scaffold region of any one of IGLV1-40 (as set out in SEQ ID NO: 18), IGLV1-44 (as set out in SEQ ID NO: 21), IGLV1-47 (as set out in SEQ ID NO: 24), IGLV1-51 (as set out in SEQ ID NO: 15), IGLV3-1 (as set out in SEQ ID NO: 6), IGLV3-19 (as set out in SEQ ID NO: 27), IGLV3-2I (as set out in SEQ ID NO: 9), IGLV6-57 (as set out in SEQ ID NO: 12); wherein the VH and the VL are capable of forming an antigen-binding site,wherein at least two of the polynucleotides differ from one another by encoding Fab fragments or derivatives thereof comprising one or more different CDRs in the VH and/or VL variable regions.49. A method of constructing a polynucleotide library , the method comprising preparing a plurality of different polynucleotides encoding a Fab fragment or derivative thereof , which comprises:a. an antibody heavy chain variable region (VH) comprising a scaffold region which is at least 90% identical to the scaffold region of IGHV3-23 as set out in SEQ ID NO: 3; and{'sub': H', ...

HIV Antigens and Antibodies

The present invention relates to a method for reducing the occurance and/or severity of viral infections. The method embodies procedures for expanding HIV from the blood of HIV antibody negative donors and deriving a non-infectious virus particle product that is antigenic. The procedures for deriving the antigenic, non-infectious virus particle product are optimally designed to maintain the integrity of the envelope proteins while maximizing the depletion of capsid proteins and RNA. The resulting virus particle product, when introduced into humans or non-human animals, enables the production of antibodies that target the natural envelope macromolecular structure that is required for infectivity. The present invention can be applied to producing virus stocks from the blood of HIV-seronegative donors, for deriving non-infectious virus particles that retain intact envelope proteins, for producing anti-viral antibodies, and for administering anti-virus antibodies to patients. 1. A composition of matter comprising isolated antigen binding proteins (ABPs) , wherein said isolated ABPs selectively bind to an epitope on an HIV-1 trimeric envelope glycoprotein subunit (TEGS).2. The composition of matter of claim 1 , wherein said TEGS is prepared by a process comprising:obtaining infectious HIV-1 virus particles from human CD4+ cell culture grown in serum-free media;contacting said infectious HIV-1 virus particles with agents that selectively remove from said particle viral RNA and viral capsid protein while retaining viral envelope protein in a non-denatured conformation, wherein said agents do not chemically fix or cross-link said envelope protein; andisolating protein from said contacted infectious HIV-1 virus particles wherein said isolated protein comprises non-infectious complexes comprising a trimeric envelope glycoprotein subunit, said subunit comprising HIV-1 envelope, gp120, and gp41 proteins that are not chemically fixed or cross-linked and substantially free of HIV ...

scFV ANTIBODY LIBRARY

The invention provides libraries of antibody molecules, libraries of nucleic acids encoding antibody molecules, methods of producing said libraries, and methods of using said libraries to select an antibody which specifically binds to an antigen. The libraries of antibody molecules include a plurality of different antibody variable domains generated by creating diversity in the CDR regions. 1. A library of antibody molecules , wherein each antibody molecule comprises (a) each solvent accessible residue in VH CDR1 and CDR2 is independently substituted with an amino acid selected from tyrosine, serine and glycine, wherein each of tyrosine, serine and glycine is equally preferred;', '(b) the VH CDR3 consists of between 8 and 17 amino acids', '(c) each solvent accessible residue in VH CDR3 is independently substituted with an amino acid selected from tyrosine, serine, glycine, alanine, phenylalanine, tryptophan, histidine, proline, valine, aspartate, asparagines, threonine and arginine in the following relative order of preference: 25% Tyr, 15% Ser, 20% Gly, 5% Ala, 5% Phe, 5% Trp, 5% His, 5% Pro, 5% Val, 3% Asp, 3% Asn, 3% Thr, 1% Arg;', '(d) the residue at position 115 of VH CDR3 is independently substituted with an amino acid selected from phenylalanine, isoleucine, leucine and methionine, wherein each of phenylalanine, isoleucine, leucine and methionine is equally preferred;, '(i) a VH domain consisting of VH CDR1, CDR2, CDR3 and framework regions, wherein the VH domain amino acid sequence is a human germline antibody heavy chain sequence in whichand (e) the VL CDR3 consists of between 8 and 12 amino acids;', '(f) each solvent accessible residue in VL CDR3 is independently substituted with an amino acid selected from tyrosine, serine, glycine, alanine, phenylalanine, tryptophan, histidine, proline, valine, aspartate, asparagines, threonine and arginine in the following relative order of preference: 25% Tyr, 15% Ser, 20% Gly, 5% Ala, 5% Phe, 5% Trp, 5% His, 5% Pro, 5 ...

SURFACE, ANCHORED FC-BAIT ANTIBODY DISPLAY SYSTEM

The present invention provides, in part, an antibody display system that simultaneously uses a secretion and a display mode. A bait complexed with a monovalent antibody fragment can be expressed on the surface of the host cell wherein the fragment may be assayed for antigen binding while full antibody is simultaneously secreted from the host cell. Methods of using the system for identifying antibodies that bind specifically to an antigen of interest are also provided. Polypeptides, polynucleotides and host cells useful for making the antibody display system are also provided along with methods of use thereof. 1. An antibody display system comprising:(a) an isolated host cell;(b) a bait comprising a heavy Fc immunoglobulin domain fused to a surface anchor polypeptide;(c) one or more polynucleotides encoding an immunoglobulin light chain variable region; and(d) one or more polynucleotides encoding an immunoglobulin heavy chain variable region.2. The antibody display system of further comprising(i) a non-tethered full antibody comprising said immunoglobulin light and heavy chains; and/or(ii) a monovalent antibody fragment which is complexed with the Fc moiety of the bait.3Pichia. The antibody display system of any one of - wherein the host cell is a cell.4. The antibody display system of any one of -wherein said one or more polynucleotides encoding an immunoglobulin light chain variable region is from a genetically diverse population of immunoglobulin light chain variable regions; and/or,wherein said one or more polynucleotides encoding an immunoglobulin heavy chain variable region is from a genetically diverse population of immunoglobulin heavy chain variable regions.5. The antibody display system of any one of - wherein the host cell comprises a polynucleotide encoding the bait which is operably associated with a regulatable promoter.6. An isolated bait polypeptide comprising an Fc immunoglobulin domain or functional fragment thereof fused claim 1 , optionally by a ...

METHODS FOR PRODUCING OPTIMISED THERAPEUTIC MOLECULES

The invention relates to a method of designing an immunoglobulin library for optimisation of a biological property of a first lead immunoglobulin and libraries of optimised immunoglobulins produced by such methods. 2. The method of claim 1 , wherein the one or more related immunoglobulins are of common lineage and bind the same target antigen as the first lead immunoglobulin claim 1 , preferably with at least 70% claim 1 , 80% claim 1 , 85% claim 1 , 90% claim 1 , 95% homology in at least one CDR region to the lead immunoglobulin.3. The method of or claim 1 , wherein the one or more related immunoglobulins have at least 70% homology in CDR3 to the lead immunoglobulin.4. The method of any preceding claim claim 1 , wherein the one or more related immunoglobulins have at least 70% homology in CDR1 and/or CDR2 to the lead immunoglobulin.5. The method of any preceding claim claim 1 , wherein the one or more related immunoglobulins have at least 70% homology in the framework regions to the lead immunoglobulin.6. The method of any preceding claim wherein the plurality of related immunoglobulins comprises at least 2 claim 1 , 3 claim 1 , 4 claim 1 , 5 claim 1 , 6 claim 1 , 7 claim 1 , 8 claim 1 , 9 claim 1 , 10 claim 1 , 15 claim 1 , 20 claim 1 , or 25 immunoglobulins.7. The method of any preceding claim wherein step c) comprises identifying sites for modification within the CDRs of the immunoglobulin sequences claim 1 , wherein a site within the CDRs is considered a site for modification if there is a variant amino acid residue present in at least one claim 1 , two claim 1 , three claim 1 , four claim 1 , or five of the related immunoglobulins.8. The method of any preceding claim wherein step c) further comprises identifying sites for modification outside the CDRs of the immunoglobulin sequences claim 1 , wherein a site outside the CDRs is considered a site for modification if there is a variant amino acid residue present in at least 20% of the related immunoglobulins.9. ...

METHOD FOR SELECTING A SINGLE CELL EXPRESSING A HETEROGENEOUS COMBINATION OF ANTIBODIES

The present invention provides combinations of specific binding proteins, such as immunoglobulins, that are designed to be true combinations, essentially all components of the combination being functional and compatible with each other. The invention further provides a method for producing a composition comprising at least two different proteinaceous molecules comprising paired variable regions, the at least two proteinaceous molecules having different binding specificities, comprising paired variable regions, at least two proteinaceous molecules having different binding specificities, comprising contacting at least three different variable regions under conditions allowing for pairing of variable regions and harvesting essentially all proteinaceous molecules having binding specificities resulting from the pairing. 143.-. (canceled)44. An expression system for producing a heterogeneous combination of monospecific and bispecific antibodies , or antibody fragments of either thereof , wherein the heterogeneous combination has specific affinity for two target epitopes , the expression system comprising:one or more nucleic acid molecules encoding three different variable regions consisting of two heavy chain variable regions and one light chain variable region together with all elements required for gene expression and paring of the variable regions;wherein antigen binding parts of the variable regions originate from an antibody from a single species;wherein one variable region is able to functionally pair with more than one other variable region; andwherein under conditions allowing for pairing of the variable regions the heterogeneous combination is produced.45. The expression system of claim 44 , wherein the one or more nucleic acid molecules encoding three different variable regions are comprised in a recombinant cell.46. The expression system of claim 45 , wherein the recombinant cell is a eukaryotic cell.47. The expression system of claim 44 , wherein the ...

METHOD FOR SELECTING A SINGLE CELL EXPRESSING A HETEROGENEOUS COMBINATION OF ANTIBODIES

The present invention provides combinations of specific binding proteins, such as immunoglobulins, that are designed to be true combinations, essentially all components of the combination being functional and compatible with each other. The invention further provides a method for producing a composition comprising at least two different proteinaceous molecules comprising paired variable regions, the at least two proteinaceous molecules having different binding specificities, comprising paired variable regions, at least two proteinaceous molecules having different binding specificities, comprising contacting at least three different variable regions under conditions allowing for pairing of variable regions and harvesting essentially all proteinaceous molecules having binding specificities resulting from the pairing. 143.-. (canceled)44. A method for producing nucleic acid sequences encoding variable regions for use in methods for producing a composition comprising at least two different proteinaceous molecules comprising paired variable regions , wherein the at least two different proteinaceous molecules have different binding specificities , the method comprising:synthesizing nucleic acid sequences encoding variable regions,expressing the nucleic acid sequences and allowing the expression products to pair, wherein the paired variable regions comprise a first variable region and a second variable region,selecting nucleic acid sequences encoding variable regions having desired pairing behavior, so as to produce nucleic acid sequences encoding variable regions.45. The method according to claim 44 , further comprising altering existing nucleic acid sequences encoding variable regions.46. The method according to claim 45 , wherein the alteration increases pairing between variable regions.47. The method of according to claim 46 , wherein a nucleic acid sequence encoding a CDR3 domain is altered.48. The method according to claim 44 , wherein the variable regions comprise ...

Synthetic library of specific binding molecules

The present invention provides methods for the production of a library of antigen specific antigen binding molecules having a peptide domain structure represented by the following formula (I): FW 1-CDR1-FW2-HV2-FW3a-HV4-FW3b-CDR3-FW4 comprising (1) isolating RNA from a member of a species in the Elasmobranchii subclass; (2) amplifying DNA sequences from RNA obtained; (3) selecting a DNA sequence from the database prepared; (4) amplifying DNA sequences encoding two or more contiguous peptide domains of FW1-CDR1-FW2-HV2-FW3a-HV4-FW3b-CDR3-FW4; (5) ligating together said amplified DNA sequences to form DNA sequences encoding an antigen specific binding molecule; (6) cloning the amplified DNA obtained into a display vector; and (7) transforming a host with said display vector to produce a library of said antigen specific antigen binding molecules. The invention also provides methods for the production of an antigen specific antigen binding molecule as defined, pharmaceutical compositions comprising such molecules and uses thereof in medicine.

METHOD FOR ISOLATION OF SOLUBLE POLYPEPTIDES

Polypeptides with biophysical properties such as solubility, stability, high expression, monomericity, binding specificity or non-aggregation, including monomeric human heavy and light chain variable domains (Vs and Vs), are identified using a high throughput method for screening polypeptides, comprising the steps of obtaining a phage display library, allowing infection of a bacterial lawn by the library phage, and identifying phage which form larger than average plaques on the bacterial lawn. Sequences of monomeric human Vs and Vs are identified, which may be useful for immunotherapy or as diagnostic agents. Multimer complexes of human Vs and Vs are also identified. The Vs and Vs identified may be used to create further libraries for identifying additional polypeptides. Further, the Vs and Vs may be subjected to DNA shuffling to select for improved biophysical properties. 1. An antigen-binding Vcomprising a FR1 sequence of EIVMTQSPGTLSLSPGDRATLSC (amino acids 1-23 of SEQ ID NO:42) , a FR2 sequence of WYQQKPGQAPRLLIY (amino acids 35-49 of SEQ ID NO:42) , a FR3 sequence of GIPDRFSGSGSGTDFTLTISRLEPEDFAVYYC (amino acids 57-88 of SEQ ID NO:42) , and a FR4 sequence of FGQGTKVTVL (amino acids 98-107 of SEQ ID NO:42).2. An antigen-binding Vcomprising the FR1 , FR2 , FR3 , and FR4 portion of SEQ ID NO:42 and one or more randomized CDR sequences , wherein one or more of CDR1 , CDR2 and CDR3 of SEQ ID NO:42 is replaced , respectively , with a randomized CDR1 , CDR2 and CDR3.3. The antigen-binding Vof claim 1 , wherein the Vis in a multimeric form.4. The antigen-binding Vof claim 1 , wherein the Vis in a dimeric form.5. The antigen-binding Vof claim 1 , wherein Vis in a trimeric form.6. The antigen-binding Vof claim 1 , wherein the Vis in a pentameric form.7. A display library constructed by preparing nucleic acid sequences coding for the antigen-binding Vof or and expressing the Vso as to display the antigen-binding Vsequence of or .8. The display library of claim 7 , wherein ...

Humanized antibodies

The present disclosure provides humanized antibodies, including antibodies comprising an ultralong CDR3 (e.g. bovine) and uses thereof.

A phage-displayed single-chain variable fragment library

Disclosed herein is a phage-displayed single-chain variable fragment (scFv) library, that comprised a plurality of phage-displayed scFvs characterized with (1) a specific CS combination; (2) a specific distribution of aromatic residues in each CDR; and (3) a specific sequence in each CDR. The present scFv library could be used to efficiently produce different antibodies with binding affinity to different antigens. Accordingly, the present disclosure provides a potential means to generate different antigen-specific antibodies promptly in accordance with the need in experimental researches and/or clinical applications.

ANTIBODIES AGAINST IMMUNOCOMPLEXES COMPRISING CYANOBACTERIAL CYCLIC PEPTIDE HEPATOTOXINS

The present invention relates to means and methods for detecting cyanobacterial cyclic peptide hepatotoxins (CCPH) in aqueous samples. More specifically, the invention provides recombinant anti-immunocomplex (anti-IC) antibodies which bind to immunocomplexes formed between one or more CCPH variants and an anti-CCPH primary antibody, and immunoassays, preferably non-competitive immunoassays, employing the same. 1. An anti-immunocomplex (anti-IC) antibody , comprising:a region configured for specifically binding to an immunocomplex formed by one or more cyanobacterial cyclic peptide hepatotoxin (CCPH) variants and an anti-Adda antibody, wherein the CCPH variant is selected from the group consisting of MC-dmRR, MC-LY, MC-LF, MC-LW, MC-WR and Nod-R.2. The anti-IC antibody according to claim 1 , comprising:a light chain variable region with CDR1 having SEQ ID NO: 2, CDR2 having SEQ ID NO: 3, and CDR3 having SEQ ID NO: 4.3. The anti-IC antibody according to claim 1 , configured for specifically recognizing an immunocomplex formed between the anti-Adda antibody and a CPPH selected from the group consisting of MC-LR claim 1 , MC-dmLR claim 1 , MC-LA claim 1 , MC-RR and MC-YR.4. The anti-IC antibody according to claim 3 , which is group-specific and configured to specifically recognize an immunocomplex formed between the anti-Adda antibody and at least CCPH variants MC-LR claim 3 , MC-dmLR claim 3 , MC-LA claim 3 , MC-RR claim 3 , MC-dmRR claim 3 , MC-YR claim 3 , MC-LY claim 3 , MC-LF claim 3 , MC-LW claim 3 , MC-WR and Nod-R claim 3 , the anti-IC antibody having a light chain variable region having CDRs 1-3 set forth in SEQ ID NO: 5 claim 3 , and a heavy chain variable region having CDRs 1-3 set forth in SEQ ID NO: 30.5. The anti-IC antibody according to claim 4 , comprising:a light chain variable region having SEQ ID NO: 5, and a heavy chain variable region having SEQ ID NO: 30.6. The anti-IC antibody according to claim 3 , which is group-specific and configured to ...

Glycan-Interacting Compounds and Methods of Use

The present invention provides glycan-interacting antibodies and methods for producing glycan-interacting antibodies useful in the treatment and prevention of human disease, including cancer. Such glycan-interacting antibodies include monoclonal antibodies, derivatives, and fragments thereof as well as compositions and kits comprising them. Further provided are methods of using glycan-interacting antibodies to target cells and treat disease.

COMPOSITIONS AND METHODS FOR GROWTH FACTOR MODULATION

Provided herein are proteins, antibodies, assays and methods useful for modulating growth factor levels and/or activities. In some embodiments, such growth factors are members of the TGF-β superfamily of proteins. 111.-. (canceled)12. An isolated monoclonal antibody , or a fragment thereof , that binds a growth factor prodomain complex (GPC) ,wherein said GPC comprises a GDF-8 growth factor and a GDF-8 prodomain (SEQ ID NO: 70),wherein the antibody or fragment thereof does not bind a mature GDF-8 growth factor, a mature GDF-11 growth factor, a GDF-11 prodomain or a complex comprising a GDF-11 prodomain and a GDF-11 growth factor, andwherein said antibody, or fragment thereof, inhibits the release of said mature GDF-8 growth factor from the GPC.13. The antibody of claim 12 , wherein said prodomain has not been cleaved at the BMP/tolloid cleavage site between residues Arg 75 and Asp 76 of the GDF-8 prodomain.14. The antibody of claim 12 , wherein said antibody is an IgG antibody.15. The antibody of claim 14 , wherein said IgG antibody is a human or humanized antibody.16. The antibody of claim 12 , wherein said antibody prevents the proteolytic cleavage by at least one metalloproteinase.17. The antibody of claim 16 , wherein said metalloproteinase is mammalian tolloid-like 2 (mTLL2).18. The antibody of claim 12 , wherein said antibody recognizes an epitope within the GDF-8 arm region of the GDF-8 prodomain set forth in SEQ ID NO: 77.19. The antibody of claim 12 , wherein said GDF-8 prodomain has not been cleaved at a furin cleavage site.20. The antibody of claim 12 , wherein said GDF-8 prodomain has been cleaved at a furin cleavage site.21. The antibody of claim 20 , wherein said furin cleavage site is Arg-Ser-Arg-Arg set forth in SEQ ID NO: 76. This application is a continuation application of U.S. application Ser. No. 14/795,012, filed on Jul. 9, 2015; which is a continuation of International Application No. PCT/US2014/036933, filed on May 6, 2014; which claims ...

ANTIBODIES AGAINST GLYPICAN-3 AND THEIR USES IN CANCER DIAGNOSIS AND TREATMENT

The present invention relates to anti-GPC3 antibodies and their applications. The invention investigates the potential inhibitory effect of anti-GPC3 antibodies on tumor growth, proliferation, migration and their applications for diagnostic and therapeutic purposes. 1. An isolated anti-GPC3 antibody or an antigen-binding portion thereof , comprising a heavy chain complementarity determining region 1 (H-CDR1) comprising the amino acid residue of SEQ ID NO: 1 , 2 , 3 or 4 , a heavy chain CDR2 (H-CDR2) comprising the amino acid residue of SEQ ID NO: 5 , 6 , 7 , 8 or 9 , and a heavy chain CDR3 (H-CDR3) comprising the amino acid residue of SEQ ID NO: 10 , 11 , 12 , 13 or 14 , anda light chain CDR1 (L-CDR1) comprising the amino acid residue of SEQ ID NO: 15, 16 or 17; a light chain CDR2 (L-CDR2) comprising the amino acid residue of SEQ ID NO: 18, 19, 20, 21 or 22, and a light chain CDR3 (L-CDR3) comprising the amino acid residue SEQ ID NO: 23, 24, 25, 26 or 27.2. (canceled)3. (canceled)4. The isolated anti-GPC3 antibody or an antigen-binding portion thereof of claim 1 , comprising:a heavy chain complementarity determining region 1 (H-CDR1) comprising the amino acid residue of SEQ ID NO: 1, a heavy chain CDR2 (H-CDR2) comprising the amino acid residue of SEQ ID NO: 5 and a heavy chain CDR3 (H-CDR3) comprising the amino acid residue of SEQ ID NO: 10; and a light chain CDR1 (L-CDR1) comprising the amino acid residue of SEQ ID NO: 15, a light chain CDR2 (L-CDR2) comprising the amino acid residue of SEQ ID NO: 18 and a light chain CDR3 (L-CDR3) comprising the amino acid residue SEQ ID NO: 23.5. (canceled)6. The isolated anti-GPC3 antibody or an antigen-binding portion thereof of claim 1 , comprising a heavy chain complementarity determining region 1 (H-CDR1) comprising the amino acid residue of SEQ ID NO: 2 claim 1 , a heavy chain CDR2 (H-CDR2) comprising the amino acid residue of SEQ ID NO: 6 and a heavy chain CDR3 (H-CDR3) comprising the amino acid residue of SEQ ID NO: 11; ...

Compositions and methods for growth factor modulation

Provided herein are proteins, antibodies, assays and methods useful for modulating growth factor levels and/or activities. In some embodiments, such growth factors are members of the TGF-β superfamily of proteins.

Conditionally active biological proteins

Methods of generating conditionally active biologic proteins, in particular therapeutic or diagnostic proteins, which are more active at an aberrant condition than at a normal physiological condition. The methods include discovery methods using libraries of proteins and assays employing physiological concentrations of components of bodily fluids. The conditionally active biologic proteins may be further evolved, conjugated to other molecules, masked, reduced in activity by attaching a cleavable moiety. Criteria for selecting starting proteins for the discovery methods, as well as formats of the proteins are also disclosed.

Methods of Sequencing, Determining, Pairing, and Validating Therapeutic Agents and Disease Specific Antigens

Provided herein are methods and composition for immune repertoire sequencing and single cell barcoding. The methods and compositions can be used to pair any two sequences originating from a single cell, such as heavy and light chain antibody sequences, for antibody discovery, disease and immune diagnostics, and low error sequencing. 1. A method comprising:(a) sequencing a polynucleotide encoding an immunoglobulin (Ig) or a T-cell receptor (TCR) polypeptide from at least one tumor infiltrating lymphocyte (TIL) from a biological sample from a subject and a polynucleotide encoding an Ig or a TCR polypeptide from at least one non-TIL cell from the biological sample from the subject, thereby obtaining sequence information; and(b) selecting an Ig or TCR polynucleotide sequence from a TIL of the at least one TIL and at least one non-TIL cell based on the sequence information.2. A method comprising:(a) sequencing a polynucleotide encoding an immunoglobulin (Ig) or a T-cell receptor (TCR) polypeptide from at least one tumor-infiltrating lymphocyte (TIL) from a biological sample from a subject and a polynucleotide encoding an Ig or a TCR polypeptide from at least one non-TIL cell from the biological sample from the subject, thereby obtaining sequence information;(b) comparing the sequence information to sequence information obtained from a corresponding normal adjacent tissue sample; and(c) selecting an Ig or TCR polynucleotide sequence from a TIL of the at least one TIL and at least one non-TIL cell based on the comparing.3. A method comprising:(a) sequencing a polynucleotide encoding an immunoglobulin (Ig) or a T-cell receptor (TCR) polypeptide from at least one tumor-infiltrating lymphocyte (TIL) from a biological sample from a first subject and a polynucleotide encoding an Ig or a TCR polypeptide from at least one non-TIL cell from the biological sample from the first subject, thereby obtaining sequence information;(b) comparing the sequence information to sequence ...

Methods and compositions with a recombinant neutralizing binding protein for treating toxin exposure

Methods, compositions and kits are provided for treating a subject exposed to or at risk for exposure to a disease agent using a pharmaceutical composition including at least one recombinant binding protein or a source of expression of the binding protein, wherein the binding protein neutralizes at least one or a plurality of disease agents that are toxins, for example at least one of a ricin toxin, a Shiga toxin, or an anthrax toxin.

Compositions and methods for growth factor modulation

Provided herein are proteins, antibodies, assays and methods useful for modulating growth factor levels and/or activities. In some embodiments, such growth factors are members of the TGF-β superfamily of proteins.

Synthetic Single Domain Antibody

The invention relates to the identification of a highly stable single domain antibody scaffold (hs2d Ab) and its use in generating synthetic single domain antibody library (hs2d Ab-L1). The invention also relates to antigen-binding proteins comprising said stable single domain antibody scaffold and their uses, in particular as therapeutics. 24-. (canceled)5. The method according to claim 1 , wherein the amino acid residues of the synthetic CDR1 and CDR2 of at least 70% claim 1 , 80% or at least 90% of the clones of the library claim 1 , are determined by the following rules:at CDR1 position 1: Y, R, S, T, F, G, A, or D;at CDR1 position 2: Y, S, T, F, G, T, or T;at CDR1 position 3: Y, S, F, or W;at CDR1 position 4: Y, R, S, T, F, G, A, W, D, E, K or N;at CDR1 position 5: S, T, F, G, A, W, D, E, N, I, H, R, Q, or L;at CDR1 position 6: S, T, Y, D, or E;at CDR1 position 7: S, T, G, A, D, E, N, I, or V;at CDR2 position 1: R, S, F, G, A, W, D, E, or Y;at CDR2 position 2: S, T, F, G, A, W, D, E, N, H, R, Q, L or Y;at CDR2 position 3: S, T, F, G, A, W, D, E, N, H, Q, P;at CDR2 position 4: G, S, T, N, or D;at CDR2 position 5: S, T, F, G, A, Y, D, E, N, I, H, R, Q, L, P, V, W, K or M;at CDR2 position 6: S, T, F, G, A, Y, D, E, N, I, H, R, Q, L, P, V, W, or K;at CDR2 position 7: S, T, F, G, A, Y, D, E, N, I, H, R, Q, L, P, or V;and wherein CDR3 amino acid sequence comprises between 9 and 18 amino acids randomly selected among one or more of the following amino acids: S, T, F, G, A, Y, D, E, N, I, H, R, Q, L, P, V, W, K, M.6. A synthetic single domain antibody library obtainable by the method of .7. The synthetic single domain antibody library of claim 6 , comprising at least 3·10distinct antibody coding sequences.89-. (canceled)10. An antigen-binding protein claim 6 , comprising a synthetic single domain antibody of the following formula: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4 claim 6 , wherein said framework regions consist of FR1 of SEQ ID NO:1 claim 6 , FR2 of SEQ ID NO:2 claim 6 , ...

ANTI-SALMONELLA ANTIBODIES AND USES THEREOF

The present disclosure provides anti-antibodies or antibody fragments, such as camelid single domain antibodies (VHHs), along with associated nucleic acids, host cells and phages. Methods of reducing the presence of in an animal or an animal environment, methods and formulations for treating infection, and methods of detecting are also described. 1. An isolated antibody or antibody fragment comprising an amino acid sequence of any one of SEQ ID NOS:1-18 , or a variant thereof.2Salmonella.. The antibody or antibody fragment of claim 1 , wherein said antibody or antibody fragment specifically binds to3Salmonella.. The antibody or antibody fragment of claim 2 , wherein said antibody or antibody fragment specifically binds to flagella of said4Salmonella. An isolated antibody or antibody fragment that binds to comprising a Complementary Determining Region (CDR)1 claim 2 , a CDR2 and a CDR3 claim 2 , wherein the CDR1 claim 2 , CDR2 and CDR3 comprise one of the following:{'sub': 1', '2', '3', '4', '5', '6', '1', '2', '3', '4', '5', '6, '(A) (i) the Complementarity Determining Region (CDR)1 comprising an amino acid sequence of GRXFSXKP (SEQ ID NO:37); (ii) the CDR2 comprising an amino acid sequence of ASXTGVST (SEQ ID NO:38); and (iii) the CDR3 comprising an amino acid sequence of AGTXRTLWGSKWRDXXEYEY (SEQ ID NO:39), wherein Xis T or S; Xis V or K; Xis F or Y; Xis T or L; Xis V or R; and Xis L or R,'}(B) (i) the CDR1 comprising an amino acid sequence of GLDFSSYA (SEQ ID NO:40), (ii) the CDR2 comprising an amino acid sequence of ISRFGGRL (SEQ ID NO:41), and (iii) the CDR3 comprising an amino acid sequence of AADRRSGLGTSKEYDY (SEQ ID NO:42),(C) (i) the CDR1 comprising an amino acid sequence of GIIFSINA (SEQ ID NO:43), (ii) the CDR2 comprising an amino acid sequence of ISAYDHT (SEQ ID NO:44), and (iii) the CDR3 comprising an amino acid sequence of NVDEIRKF (SEQ ID NO:45),(D) (i) the CDR1 comprising an amino acid sequence of GRSFSLYG (SEQ ID NO:46), (ii) the CDR2 comprising an ...

METHOD FOR SELECTING A SINGLE CELL EXPRESSING A HETEROGENEOUS COMBINATION OF ANTIBODIES

Described are combinations of specific binding proteins, such as immunoglobulins, that are designed to be true combinations, essentially all components of the combination being functional and compatible with each other. Further provided are methods for producing a composition comprising at least two different proteinaceous molecules comprising paired variable regions, the at least two proteinaceous molecules having different binding specificities, comprising paired variable regions, at least two proteinaceous molecules having different binding specificities, comprising contacting at least three different variable regions under conditions allowing for pairing of variable regions and harvesting essentially all proteinaceous molecules having binding specificities resulting from the pairing. 1. A method for selecting a single recombinant cell that expresses a heterogeneous combination of monospecific and bispecific antibodies , or antibody fragments thereof , wherein the monospecific and bispecific antibodies , or antibody fragments thereof , are human , humanized , or deimmunized , and wherein the heterogeneous combination has specific affinity for two target epitopes , the method comprising:carrying out a process for producing the heterogeneous combination; the process comprising providing three different variable regions consisting of two heavy chain regions and one light chain variable region in recombinant cells, wherein antigen binding parts of the variable regions originate from a single species and wherein one variable region is able to functionally pair with more than one other variable region, and under conditions allowing for pairing of variable regions and secretion of the paired regions from the recombinant cells resulting in the production of said heterogeneous combination,providing two target epitopes, andselecting a single recombinant cell from the recombinant cells that produces a heterogeneous combination that binds the two target epitopes,wherein the ...

Single domain serum albumin binding protein

Disclosed herein are single domain serum albumins binding proteins with improved thermal stability, binding affinities, and robust aggregation profiles. Also described are multispecific binding proteins comprising a single domain serum albumin binding protein according to the instant disclosure. Pharmaceutical compositions comprising the binding proteins disclosed herein and methods of using such formulations are provided.

ANTI-FACTOR IX PADUA ANTIBODIES

Provided herein are anti-Factor IX Padua binding constructs, e.g., antibodies and antigen-binding fragments thereof. Related polypeptides, conjugates and kits are also provided. The inventions may be used in methods of detecting Factor IX Padua in a sample. 139.-. (canceled)40. An anti-Factor IX Padua binding construct comprising two or more antigen-binding fragments linked together , wherein at least one of the antigen-binding fragments comprises the amino acid sequences of: SSYAIS (SEQ ID NO: 6) , GIVPAFGTANYAQKFQG (SEQ ID NO: 7) , SWGVISFAY (SEQ ID NO: 8) , RASQDISSYLN (SEQ ID NO: 9) , AASNLQS (SEQ ID NO: 10) , and MQYDSLPFTF (SEQ ID NO: 11).41. The anti-Factor IX Padua binding construct of claim 40 , wherein at least one of the antigen-binding fragments comprises the amino acid sequences of SEQ ID NOs: 24 and 25 or SEQ ID NOs: 26 and 27.42. The anti-Factor IX Padua binding construct of claim 40 , wherein the two or more antigen-binding fragments are linked together via a disulfide bond claim 40 , a helix-turn-helix structure claim 40 , or an alkaline phosphatase domain.43. The anti-Factor IX Padua binding construct of claim 40 , wherein at least one of the antigen-binding fragments is a Fab antibody fragment claim 40 , a Fab2′ antibody fragment claim 40 , or a Fab mini antibody.44. The anti-Factor IX Padua binding construct of claim 40 , further comprising a FLAG tag comprising DYKDDDDK (SEQ ID NO: 12) and/or a hexa-His tag comprising HHHHHH (SEQ ID NO: 13) claim 40 , optionally claim 40 , wherein the FLAG tag and/or the hexa-His tag are located at the C-terminal end of the anti-Factor IX Padua binding construct.45. The anti-Factor IX Padua binding construct of claim 40 , wherein the anti-Factor IX Padua binding construct is a dimerized Fab2′ antibody fragment linked via an alkaline phosphatase domain.46. A nucleic acid comprising a nucleotide sequence encoding the anti-Factor IX Padua binding construct of .47. A vector comprising the nucleic acid of .48. A host ...

SURFACE, ANCHORED FC-BAIT ANTIBODY DISPLAY SYSTEM

The present invention provides, in part, an antibody display system that simultaneously uses a secretion and a display mode. A bait complexed with a monovalent antibody fragment can be expressed on the surface of the host cell wherein the fragment may be assayed for antigen binding while full antibody is simultaneously secreted from the host cell. Methods of using the system for identifying antibodies that bind specifically to an antigen of interest are also provided. Polypeptides, polynucleotides and host cells useful for making the antibody display system are also provided along with methods of use thereof. 120-. (canceled)21. A method for making an antibody display system comprising:(a) an isolated eukaryotic host cell;(b) a bait comprising a human Fc immunoglobulin domain fused to a surface anchor polypeptide;(c) one or more polynucleotides encoding an immunoglobulin light chain variable region;(d) one or more polynucleotides encoding an immunoglobulin heavy chain variable region;comprising introducing, into said eukaryotic host cell, a polynucleotide encoding said bait, said one or more polynucleotides encoding an immunoglobulin light chain variable region; and said one or more polynucleotides encoding an immunoglobulin heavy chain variable region.22. A method for making an antibody or antigen-binding fragment thereof comprisingintroducing, into an isolated eukaryotic host cell comprising a bait that includes a human Fc immunoglobulin domain or functional fragment thereof fused to a surface anchor polypeptide or functional fragment thereof, one or more polynucleotides encoding an immunoglobulin light chain variable region; and/or one or more polynucleotides encoding an immunoglobulin heavy chain variable region; andculturing the host cell under condition whereby the polynucleotides encoding the immunoglobulin chains are expressed and an antibody or antigen-binding fragment thereof is formed from said chains;wherein said bait is operably associated with a ...

Method for producing antibody

Disclosed is an antibody in which the 80th amino acid residue in a variable region based on the Kabat method and the 171th amino acid residue in a constant region based on the Kabat method are substituted with cysteine in an antibody in which the 80th amino acid residue in the variable region and the 171th amino acid residue in the constant region are not cysteine.

Bispecific single chain antibodies with specificity for high molecular weight target antigens

The present invention provides a method for the selection of bispecific single chain antibodies comprising a first binding domain capable of binding to an epitope of CD3 and a second binding domain capable of binding to the extracellular domain cell surface antigens with a high molecular weight extracellular domain. Moreover, the invention provides bispecific single chain antibodies produced by the use of the method of the invention, nucleic acid molecules encoding these antibodies, vectors comprising such nucleic acid molecules and methods for the production of the antibodies. Furthermore, the invention provides pharmaceutical compositions comprising bispecific single chain antibodies of the invention, medical uses of the same and methods for the treatment of diseases comprising the administration of bispecific single chain antibodies of the invention.

Polypeptides, antibody variable domains & antagonists

The invention relates to anti-TNFR1 polypeptides and antibody single variable domains (dAbs) that are resistant to degradation by a protease, as well as antagonists comprising these. The polypeptides, dAbs and antagonists are useful for as therapeutics and/or prophylactics that are likely to encounter proteases when administered to a patient, for example for pulmonary administration, oral administration, delivery to the lung and delivery to the GI tract of a patient, as well as for treating inflammatory disease, such as arthritis or COPD.

Glycan-Interacting Compounds and Methods of Use

The present invention provides glycan-interacting antibodies and methods for producing glycan-interacting antibodies useful in the treatment and prevention of human disease, including cancer. Such glycan-interacting antibodies include monoclonal antibodies, derivatives, and fragments thereof as well as compositions and kits comprising them. Further provided are methods of using glycan-interacting antibodies to target cells and treat disease.

ANTI-SALMONELLA ANTIBODIES AND USES THEREOF

The present disclosure provides anti-antibodies or antibody fragments such as camelid single domain antibodies (VHHs), along with associated nucleic acids, host cells and phages. Methods of reducing the presence of in an animal or an animal environment, methods and formulations for treating infection, and methods of detecting are also described. 1Salmonella(i) the CDR1 comprising an amino acid sequence of GLDFSSYA (SEQ ID NO:40),(ii) the CDR2 comprising an amino acid sequence of ISRFGGRL (SEQ ID NO:41), and(iii) the CDR3 comprising an amino acid sequence of AADRRSGLGTSKEYDY (SEQ ID NO:42).. An isolated antibody or antibody fragment that binds to comprising a Complementary Determining Region (CDR)1, a CDR2 and a CDR3, wherein the CDR1, CDR2 and CDR3 comprise the following: The field of the present invention relates generally to antibodies, fragments thereof, derivatives thereof, and to uses and applications of such antibodies. The antibodies and fragments described may be specifically directed againstSalmonellosis is one of the most commonly reported zoonotic diseases in humans. In the United States alone, it causes an estimated 1.3 million human food-borne illnesses and more than 500 deaths each year (Messens et al., 2013). serotypes and are frequently detected in human infections (Ravel et al., 2010). Salmonellas are widely distributed in nature, and they are commonly carried by wild or farm-animal vectors. Poultry is known to be a major global reservoir of Salmonellas. live in poultry gut as transient members of the intestinal microbial population without causing disease. Colonization of does not usually affect poultry body weight gain or performance; thus, asymptomatic infection can increase the likelihood of zoonotic transmission to humans through the food chain (Hugas et al., 2014; Mazengia et al., 2014). Chicks can become infected vertically (from adults via the egg to the chick) or horizontally (from the environment, pests, or feed) (Cox et al., 2014; ...

ION CONCENTRATION-DEPENDENT BINDING MOLECULE LIBRARY

Disclosed is a library consisting essentially of a plurality of antigen-binding molecules differing in sequence from each other, wherein an antigen-binding domain in each of the antigen-binding molecules comprises at least one amino acid residue that changes the antigen-binding activity of the antigen-binding molecule depending on ion concentration conditions. Also disclosed are a composition comprising a plurality of polynucleotide molecules each encoding the antigen-binding molecules, a composition comprising a plurality of vectors each comprising the polynucleotide molecules, a method for selecting the antigen-binding molecules, a method for isolating the polynucleotide molecules, a method for producing the antigen-binding molecules, and a pharmaceutical composition comprising any of the antigen-binding molecules. 122.-. (canceled)23. A library consisting essentially of a plurality of antigen-binding molecules differing in sequence from each other , wherein an antigen-binding domain in each of the antigen-binding molecules comprises at least one amino acid residue that changes the antigen-binding activity of the antigen-binding molecule depending on ion concentration conditions , and wherein the ion concentration conditions are pH conditions.24. The library according to claim 23 , wherein the amino acid residue is contained in the antigen-binding domain in a heavy chain of the antigen-binding molecule.25. The library according to claim 24 , wherein the antigen-binding domain in a heavy chain is a heavy chain variable region.26. The library according to claim 25 , wherein the amino acid residue is located at any one or more of positions 27 claim 25 , 31 claim 25 , 32 claim 25 , 33 claim 25 , 35 claim 25 , 50 claim 25 , 52 claim 25 , 53 claim 25 , 55 claim 25 , 57 claim 25 , 58 claim 25 , 59 claim 25 , 61 claim 25 , 62 claim 25 , 95 claim 25 , 96 claim 25 , 97 claim 25 , 98 claim 25 , 99 claim 25 , 100a claim 25 , 100b claim 25 , 100d claim 25 , 100f claim 25 , ...

ENGINEERING OF IMMUNOGLOBULIN DOMAINS

The present invention provides a single domain antibody (sdAb) scaffold comprising one or more than one non-canonical disulfide bond in the framework region (FR). The one or more than one non-canonical disulfide bond may be formed between cysteines introduced by mutations in FR2 and FR3. In the case where the sdAb scaffold is a V, the Cys may be introduced at any one of positions 47-49 and any one of positions 67-71, based on Kabat numbering; in one example, the Cys may be introduced at positions 49 and 69, based on Kabat numbering. In the case where the sdAb scaffold is a V, the Cys residues may be introduced at any one of positions 46-49 and any one of positions 62-66, based on Kabat numbering; in one example, the Cys residues may be introduced at positions 48 and 64, based on Kabat numbering. 1. A synthetic immunoglobulin scaffold comprising one or more than one non-canonical disulfide bond in the framework region (FR) , wherein the immunoglobulin scaffold is a human Vscaffold , and wherein the non-canonical disulfide bond is formed between cysteine residues introduced at positions 48 and 64 , based on Kabat numbering.2. The synthetic immunoglobulin scaffold of claim 1 , wherein the synthetic immunoglobulin scaffold is immobilized onto a surface.3. A polypeptide comprising the synthetic immunoglobulin scaffold of claim 1 , selected from the group consisting of a single domain antibody (sdAb) claim 1 , scFv claim 1 , Fab claim 1 , F(ab) claim 1 , and mature immunoglobulin.4. The polypeptide of claim 3 , wherein the mature immunoglobulin is selected from the group consisting of an IgG1 claim 3 , IgG2 claim 3 , IgG3 claim 3 , IgG4 claim 3 , IgE claim 3 , and/or IgM.5. A composition comprising the synthetic immunoglobulin scaffold of .6. (canceled)7. (canceled)8. The synthetic immunoglobulin scaffold of wherein the Vis of the kappa or lambda family.911.-. (canceled)12. The polypeptide of claim 3 , wherein the synthetic immunoglobulin scaffolds are multimerized claim ...

PHAGE-DISPLAYED ANTIBODY LIBRARIES AND USES THEREOF

Disclosed herein are phage-displayed single-chain variable fragment (scFv) libraries, which comprised a plurality of scFvs with a specific sequence in each CDR. The present scFv libraries could be used to efficiently produce different antibodies with high binding affinity to H1 hemagglutinin of influenza virus. Accordingly, the present disclosure provides a potential means to generate different antigen-specific antibodies promptly in accordance with the need in experimental researches and/or clinical applications. 1. A phage-displayed single-chain variable fragment (scFv) library comprising a plurality of scFvs that are expressed by a phage and exhibit binding affinity and specificity to a protein antigen , wherein each of the plurality of scFvs comprises at least one polypeptide selected from the group consisting of ,a first complementarity determining region (CDR) encoded by the nucleotide sequence of SEQ ID NO: 1,a second CDR encoded by the nucleotide sequence of SEQ ID NO: 2, anda third CDR encoded by a the nucleotide sequence of SEQ ID NO: 3.2. The phage-displayed scFv library of claim 1 , wherein the phage is a M13 phage or a T7 phage.3. The phage-displayed scFv library of claim 1 , wherein the protein antigen is a hemagglutinin of influenza virus.4. A method for selecting one phage-displayed scFv from the phage-displayed scFv library of claim 1 , wherein the phage-displayed scFv exhibits high affinity and specificity to a protein antigen claim 1 , comprising claim 1 ,{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, '(a) incubating the phage-displayed scFv library of with protein antigens, wherein the phage-displayed scFv library comprises a plurality of phage-displayed scFvs;'}(b) subjecting the product of step (a) to an acid treatment thereby producing a plurality of phage-displayed scFvs, which are respectively bound to the protein antigens before the acid treatment;(c) subjecting the plurality of phage-displayed scFvs produced in the step (b) to an ...

Method for recovering two or more genes, or gene products, encoding an immunoreceptor

The present invention is related to a method for recovering two or more genes, or gene products, or cDNAs, encoding for an immunoreceptor having two or more subunits, which two or more genes, or gene products, are comprised in a given source cell. The invention is further related to a method of creating a library of expressor cells, in which library each cell is capable of expressing two or more genes, or gene products, encoding for the subunits of the immunoreceptor. The invention is further related to a method of screening a library of expressor cells as created according to the above method, for one cell that expresses an immunoreceptor that has specificity for a given target molecule.

Recombinant polynucleotide and a transgenic flammulina velutipes carrying the same

The invention provides a recombinant polynucleotide comprising a truncated glyceraldehyde-3-phosphate dehydrogenase (gpd) promoter and a modified HBV S protein gene and a transgenic Flammulina velutipes carrying the recombinant polynucleotide. The invention surprisingly found that after administering the transgenic Flammulina velutipes to a subject, the subject can successfully generate an antibody against HBV. Therefore, the transgenic Flammulina velutipes can be used as a vaccine against HBV.

METHOD FOR ISOLATION OF SOLUBLE POLYPEPTIDES

Polypeptides with desirable biophysical properties such as solubility, stability, high expression, monomericity, binding specificity or non-aggregation, including monomeric human Vs and Vs, are identified using a high throughput method for screening polypeptides, comprising the steps of obtaining a phage display library, allowing infection of a bacterial lawn by the library phage, and identifying phage which form larger than average plaques on the bacterial lawn. Sequences of monomeric human Vs and Vs are identified, which may be useful for immunotherapy or as diagnostic agents. Multimer complexes of human Vs and Vs are also identified. The Vs and Vs identified may be used to create further libraries for identifying additional polypeptides. Further, the Vs and Vs may be subjected to DNA shuffling to select for improved biophysical properties. 1. An antibody fragment comprising a FR1 sequence of QLQLQESGGGLVQPGGSLRLSCAASGFTFS (amino acids 1-30 of SEQ ID NO:16) , a FR2 sequence of WFRQAPGKGLEWVG (amino acids 36-49 of SEQ ID NO:16) , a FR3 sequence of RFTISRDDSKSIAYLQMNSLRAEDTAMYYCAR (amino acids 69-100 of SEQ ID NO:16) , and a FR4 sequence of WGQGTLVTVSS (amino acids 113-123 of SEQ ID NO:16).2. An antibody fragment comprising the FR1 , FR2 , FR3 , and FR4 portion of SEQ ID NO:16.3100. The antibody fragment of claim , wherein the antibody fragment is in a multimeric form.4100. The antibody fragment of claim , wherein the antibody fragment is in a dimeric form.5100. The antibody fragment of claim , wherein the antibody fragment is in a trimeric form.6100. The antibody fragment of claim , wherein the antibody fragment is in a pentameric form.7100101. A display library constructed comprising the antibody fragment sequence of claim or .8106. The display library of claim , wherein the library is a phage display library.9106. The display library of claim , wherein the library is a ribosome display , ARM ribosome display , yeast display , bacterial cell display , or in vitro ...

NOVEL MULTIVALENT IMMUNOGLOBULINS

The present invention provides a multivalent immunoglobulin or part thereof binding specifically to at least two cell surface molecules of a single cell with at least one modification in at least one structural loop region of said immunoglobulin determining binding to an epitope of said cell surface molecules wherein the unmodified immunoglobulin does not significantly bind to said epitope, its use and methods for producing it. 1. A polypeptide comprising a scaffold comprising an immunoglobulin fold of an antibody CH3 constant domain , wherein(i) said antibody CH3 constant domain comprises at least six structural loops comprising up to 30 amino acid residue changes relative to the corresponding amino acid residues of said antibody CH3 constant domain; and(ii) one or more solvent accessible surfaces are formed by at least three of said residue changes in two of said structural loops or by at least four of said residue changes in one of said structural loops.2. The polypeptide of claim 1 , wherein at least two solvent accessible surfaces are formed by said residue changes.3. The polypeptide of claim 1 , wherein said polypeptide comprises an antibody variable domain.4. The polypeptide of claim 1 , wherein said immunoglobulin fold is of a human antibody CH3 constant domain.5. The polypeptide of claim 1 , wherein said immunoglobulin fold is of a antibody CH3 constant domain having an isotype selected from the group consisting of IgG and IgA.6. The polypeptide of claim 5 , wherein said IgG isotype is human and is selected from the group consisting of IgG1 claim 5 , IgG2 claim 5 , IgG3 and IgG4.7. The polypeptide of claim 1 , wherein said six structural loops of (i) comprise the AB loop claim 1 , the CD loop and the EF loop claim 1 , and said two structural loops of (ii) or said one structural loop of (ii) are loop(s) selected from the group consisting of the AB loop claim 1 , the CD loop and the EF loop.8. The polypeptide of claim 7 , wherein said immunoglobulin fold is ...

Antigen-binding domain, and polypeptide including conveying section

The present invention relates to a polypeptide comprising an antigen binding domain and a carrying moiety having an inhibiting domain that inhibits the antigen binding activity of the antigen binding domain, and having a longer half-life than that of the antigen binding domain existing alone, methods for producing and screening for the polypeptide, a pharmaceutical composition comprising the polypeptide, methods for producing and screening for a single-domain antibody whose antigen binding activity is inhibited by associating with particular VL, VH or VHH, and a fusion polypeptide library including a single-domain antibody whose antigen binding activity is inhibited by associating with particular VL, VH or VHH.

ANTI-VEGF ANTIBODIES

Anti-VEGF antibodies and variants thereof, including those having high affinity for binding to VEGF, are disclosed. Also provided are methods of using phage display technology with naïve libraries to generate and select the anti-VEGF antibodies with desired binding and other biological activities. Further contemplated are uses of the antibodies in research, diagnostic and therapeutic applications. 120-. (canceled)21. An isolated nucleic acid molecule encoding an antibody , or an antibody fragment thereof , wherein the antibody , or antibody fragment thereof , binds to human VEGF and comprises a heavy chain variable domain (VH) and a light chain variable domain (VL) , (a) CDR-H1 comprises the amino acid sequence DYWIH (SEQ ID NO: 261),', '(b) CDR-H2 comprises the amino acid sequence GVTPAGGYTYYADSVKG (SEQ ID NO: 944), and', '(c) CDR-H3 comprises the amino acid sequence FVFFLPYAMDY (SEQ ID NO: 475); or', '(a) CDR-H1 comprises the amino acid sequence DYWIH (SEQ ID NO: 261),', '(b) CDR-H2 comprises the amino acid sequence GVTPAGGYTAYADSVKG (SEQ ID NO: 945), and', '(c) CDR-H3 comprises the amino acid sequence FVFFLPYAMDY (SEQ ID NO: 475);, 'wherein the VH comprises a CDR-H1, CDR-H2, and CDR-H3, wherein (d) CDR-L1 comprises the amino acid sequence RASQDVSTAVA (SEQ ID NO: 48),', '(e) CDR-L2 comprises the amino acid sequence SASFLYS (SEQ ID NO: 49), and', '(f) CDR-L3 comprises the amino acid sequence KQGFANPFT (SEQ ID NO: 946)., 'and wherein the VL comprises a CDR-L1, CDR-L2, and CDR-L3, wherein24. A vector comprising the nucleic acid of .25. A host cell comprising the vector of .26. A process of producing an antibody claim 25 , or antibody fragment thereof claim 25 , comprising culturing the host cell of so that the antibody is produced.27. The process of claim 26 , wherein the antibody claim 26 , or antibody fragment thereof claim 26 , is recovered from the host cell medium. This application is a continuation of U.S. patent application Ser. No. 14/636,782, filed on Mar. 3 ...

Preparation of libraries of protein variants expressed in eukaryotic cells and use for selecting binding molecules

The invention relates to methods of producing eukaryotic cell libraries encoding a repertoire of binding molecules (“binders”), wherein the methods use a site-specific nuclease for targeted cleavage of cellular DNA to enhance site-specific integration of binder genes through endogenous cellular repair mechanisms. Populations of eukaryotic cells are produced in which a repertoire of genes encoding binders are integrated into a desired locus in cellular DNA (e.g., a genomic locus) allowing expression of the encoded binding molecule, thereby creating a population of cells expressing different binders.

Immunoglobulin Variable Region Libraries

Antigen-specific immunoglobulin V-regions are identified from a library of nucleic acids amplified using polymerase chain reaction using leader sequence-specific forward primers. The use of leader sequence primers allows all V-region sequences to be amplified (including those with extensive 5′ end mutations) without loss of the original 5′ V gene segment sequence. These libraries can be screened for antigen-specific V-regions using eukaryotic cells engineered to express the amplified V-region-encoding nucleic acids or using bacterial phage display techniques. In the latter, a second V-region library is made using a larger than conventional set of 5′ V-region primers. The sequence errors introduced into the amplification products by this method are corrected using sequence information obtained in the products amplified by the V-region primers to screen the library created using the leader sequence primers. Amino acid sequence information from fragments of donor immunoglobulins can be used to assist in the identification of nucleic acids encoding the heavy and light chains of donor antibodies as well as to design primers to amplify such nucleic acids. 1. A purified monoclonal antibody comprising a variable region amino acid sequence that promotes binding of the antibody to a target antigen and has somatic hypermutations in its first N-terminal 8 amino acids , wherein the variable region amino acid sequence is determined by screening a library of human immunoglobulin nucleotide sequences made by a method comprising the steps of:(a) using a first PCR to amplify human immunoglobulin heavy and light chain nucleotide sequences from a cDNA library made from a sample of human peripheral blood mononuclear cells obtained from a human donor previously determined to possess antibodies specific for target antigens of interest, wherein leader-specific forward primers that amplify at least 90% of the human immunoglobulin V-region genes and reverse primers for amplifying nucleic ...

MOLECULES WITH ALTERED NEONATE FC RECEPTOR BINDING HAVING ENHANCED THERAPEUTIC AND DIAGNOSTIC PROPERTIES

The present invention provides molecules, including proteins, more particularly, immunoglobulins whose in vivo half-lives are altered (increased or decreased) by the presence of an IgG constant domain, or FcRn binding fragment thereof (e.g., an Fc region or hinge-Fc region) (e.g., from a human IgG, e.g., human IgG1), that have modifications of one or more of amino acid residues in at least the CH3 domain. 1. A modified human or humanized IgG1 comprising an Fc region comprising amino acid substitutions at two or more of positions 432 to 437 , numbered according to the EU numbering index of Kabat , relative to a human wild-type Fc region; wherein(i) positions 432 and 437 are each substituted with cysteine;(ii) position 433 is histidine or is substituted with arginine, proline, threonine, lysine, serine, alanine, methionine, or asparagine;(iii) position 434 is asparagine or is substituted with arginine, tryptophan, histidine, phenylalanine, tyrosine, serine, methionine or threonine;(iv) position 435 is histidine or is substituted with histidine; and(v) position 436 is tyrosine or phenylalanine or is substituted with leucine, arginine, isoleucine, lysine, methionine, valine, histidine, serine, or threonine;and wherein the modified human or humanized IgG1 has an increased half-life compared to the half-life of an IgG1 having the human wild-type Fc region.2. (canceled)3. (canceled)4. The modified human or humanized IgG1 of claim 1 , further comprising an amino acid insertion after position 437.5. The modified human or humanized IgG1 of claim 4 , wherein the amino acid insertion is glutamic acid.6. The modified human or humanized IgG1 of claim 1 , wherein the binding affinity of the modified human or humanized IgG1 for FcRn at pH 6.0 is higher than the binding affinity of the IgG1 having the human wild-type Fc region for FcRn at pH 6.7. The modified human or humanized IgG1 of claim 1 , wherein the binding affinity of the modified human or humanized IgG1 for FcRn at pH 7.4 ...

RADICALLY DIVERSE HUMAN ANTIBODY LIBRARY

Disclosed is an antibody library comprising a plurality of antibodies with non-naturally occurring combinations of complementary determining regions from memory and naïve B-cells naturally occurring in humans, and wherein the antibody library comprises a high number of functional and non-redundant antibodies. Further disclosed are methods of preparing antibody libraries with a high level of functional diversity. 1. An antibody library that comprises a plurality of antibodies , a) a VH domain that comprises a VH-CDR1 sequence, a VH-CDR2 sequence, a VH-CDR3 sequence; and', 'b) a VL domain that comprises a VL-CDR1 sequence, a VL-CDR2 sequence, a VL-CDR3 sequence; and, 'wherein each antibody of the plurality of antibodies comprises a) at least one of the VH-CDR3 sequence and the VL-CDR3 sequence is derived from a naïve B-cell;', 'b) if only one of the VH-CDR3 sequence and the VL-CDR3 sequence is derived from the naïve B-cell, then the VH-CDR3 sequence or VL-CDR3 sequence not derived from the naïve B-cell is derived from a memory cell; and', 'c) the VH-CDR1 sequence, VH-CDR2 sequence, VL-CDR1 sequence, and VL-CDR2 sequence are derived from a memory B-cell., 'wherein2. The antibody library of claim 1 , wherein the at least one of the VH-CDR3 sequence and the VL-CDR3 sequence derived from a naïve B-cell is a naturally occurring sequence.3. The antibody library of claim 1 , wherein the VH-CDR3 sequence or VL-CDR3 sequence derived from a memory cell is a naturally occurring sequence.4. The antibody library of claim 1 , wherein the VH-CDR1 sequence claim 1 , VH-CDR2 sequence claim 1 , VL-CDR1 sequence claim 1 , and VL-CDR2 sequence derived from a memory B cell are naturally occurring sequences.5. (canceled)6. (canceled)7. (canceled)8. The antibody library of claim 1 , wherein the VL domain is a Vκ domain or a Vλ domain.9. The antibody library of claim 1 , wherein the naïve B-cell is a CD27−/IgM+ B-cell or a CD27−/IgD+ B-cell.10. The antibody library of claim 1 , wherein the ...

Methods of Sequencing, Determining, Pairing, and Validating Therapeutic Agents and Disease Specific Antigens

Provided herein are methods and composition for immune repertoire sequencing and single cell barcoding. The methods and compositions can be used to pair any two sequences originating from a single cell, such as heavy and light chain antibody sequences, for antibody discovery, disease and immune diagnostics, and low error sequencing. 1255-. (canceled)256. An immunoglobulin (Ig) or T-cell receptor (TCR) polypeptide produced by a process comprising: a single tumor infiltrating lymphocyte (TIL) cell or a single non-TIL cell, wherein the single cell is isolated from a first biological sample from a first subject,', 'a molecular barcoded polynucleotide,', 'a vessel barcoded polynucleotide,', 'a forward primer and a reverse primer for amplifying the vessel barcoded polynucleotide, and', 'a reverse transcriptase,, '(a) forming a plurality of vessels, at least one vessel comprisingwherein the molecular barcoded polynucleotide comprises a 5′ region complementary to a region of the vessel barcoded polynucleotide;(b) generating a cDNA polynucleotide by reverse transcription of an RNA from the single cell, wherein the reverse transcriptase adds three or more non-template nucleotides to the 3′ end of the cDNA polynucleotide;(c) annealing the molecular barcoded polynucleotide to the three or more non-template nucleotides of the cDNA polynucleotide, and extending the cDNA polynucleotide to generate a single-barcoded cDNA polynucleotide;(d) amplifying the vessel barcoded polynucleotide using the forward primer and the reverse primer, thereby generating an amplified product, annealing the amplified product to the single-barcoded cDNA polynucleotide, and extending the single-barcoded cDNA polynucleotide to generate a dual-barcoded cDNA polynucleotide;(e) sequencing the dual-barcoded cDNA polynucleotide thereby obtaining sequence information;(f) selecting an Ig or a TCR polynucleotide sequence from a TIL based on the sequence information;(g) producing an Ig or TCR polypeptide encoded ...

METHOD OF IMPROVING CHARACTERISTICS OF PROTEINS

The invention provides efficient methods for combining single-substitution libraries of nucleic acids that span and encode proteins of interest and for selecting resultant mutant proteins after expression which have improved properties or characteristics. Specifically, the methods comprising synthesizing a single substitution library for each of a plurality of domains of a protein; expressing separately each member of each single substitution library as a pre-candidate protein; selecting members of each single substitution library which encode pre-candidate proteins which exhibit an improvement in the one or more predetermined characteristics to form a selected library; shuffling members or the selected libraries in a PCR to produce a combinatorial shuffled library; expressing members of the shuffled library as candidate proteins; and selecting mutant proteins which have improved properties or characteristics. 1. A method of improving one or more predetermined characteristics of a protein , the method comprising the steps of:synthesizing a single substitution library for each of a plurality of domains of a protein, each member of a single substitution library having a nucleotide sequence that overlaps a nucleotide sequence of at least one member of a different single substitution library;expressing separately each member of each single substitution library as a pre-candidate protein;selecting members of each single substitution library which encode pre-candidate proteins which exhibit an improvement in the one or more predetermined characteristics to form a selected library for each domain of the protein;shuffling members of the selected libraries in a PCR to produce a combinatorial shuffled library;expressing members of the shuffled library as candidate proteins; andselecting members of the shuffled library which encode candidate proteins which exhibit an improvement in at least one of the one or more predetermined characteristics.2. The method of wherein said ...

ANTI-LIPOARABINOMANNAN ANTIBODY AND IMMUNOASSAY FOR ACID-FAST BACILLARY INFECTION USING THE ANTIBODY

The present invention provides a monoclonal antibody that specifically binds to acid-fast bacillary lipoarabinomannan, particularly tubercle bacillary lipoarabinomannan, the antibody being set forth below: 1. A monoclonal antibody that is capable of binding to acid-fast bacillary lipoarabinomannan , the antibody being set forth in any one of (A) to (C) below: (a) a heavy chain CDR1 consisting of the amino acid sequence set forth in SEQ ID NO: 1,', '(b) a heavy chain CDR2 consisting of the amino acid sequence set forth in SEQ ID NO: 2,', '(c) a heavy chain CDR3 consisting of the amino acid sequence set forth in SEQ ID NO: 3,', '(d) a light chain CDR1 consisting of the amino acid sequence set forth in SEQ ID NO: 4,', '(e) a light chain CDR2 consisting of the amino acid sequence set forth in SEQ ID NO: 5, and', '(f) a light chain CDR3 consisting of the amino acid sequence set forth in SEQ ID NO: 6;, '(A) a monoclonal antibody comprising a heavy chain variable region and a light chain variable region joined via a linker, the heavy chain variable region comprising heavy chains CDR1 to CDR3 shown in (a) to (c) below, and the light chains comprising CDR1 to CDR3 shown in (d) to (f) below (g) a heavy chain CDR1 consisting of the amino acid sequence set forth in SEQ ID NO: 31,', '(h) a heavy chain CDR2 consisting of the amino acid sequence set forth in SEQ ID NO: 32,', '(i) a heavy chain CDR3 consisting of the amino acid sequence set forth in SEQ ID NO: 33,', '(j) a light chain CDR1 consisting of the amino acid sequence set forth in SEQ ID NO: 34,', '(k) a light chain CDR2 consisting of the amino acid sequence set forth in SEQ ID NO: 35, and', '(l) a light chain CDR3 consisting of the amino acid sequence set forth in SEQ ID NO: 36; and, '(B) a monoclonal antibody comprising a heavy chain variable region and a light chain variable region joined via a linker, the heavy chain variable region comprising heavy chains CDR1 to CDR3 shown in (g) to (i) below, and the light chains ...

ENGINEERED ANTIBODY CONSTANT DOMAIN MOLECULES

Described herein are engineered antibody constant domain molecules, such as CH2 or CH3 domain molecules, comprising at least one mutation, or comprising at least one complementarity determining region (CDR), or a functional fragment thereof, engrafted in a loop region of the CH2 domain. The CH2 domain molecules described herein are small, stable, soluble, exhibit little to no toxicity and are capable of binding antigen. 1. A polypeptide comprising an immunoglobulin CH2 domain of IgG , IgA or IgD , or a CH3 domain of IgE or IgM , wherein the CH2 domain or CH3 domain comprises an N-terminal truncation of about 1 to about 7 amino acids , and wherein (i) at least one of the loops of the CH2 or CH3 domain is mutated; (ii) at least a portion of a loop region of the CH2 domain or CH3 domain is replaced by a complementarity determining region (CDR) , or a functional fragment thereof , from a heterologous immunoglobulin variable domain; or (iii) both , wherein the polypeptide has a molecular weight of less than about 15 kD , and wherein the polypeptide specifically binds an antigen.2. The polypeptide of claim 1 , wherein Loop 1 of the CH2 or CH3 domain is mutated.3. The polypeptide of claim 2 , wherein the CH2 domain or CH3 domain further comprises a mutated Loop 2 claim 2 , a mutated Loop 3 claim 2 , a mutated Loop A-B claim 2 , a mutated Loop C-D claim 2 , a mutated Loop E-F claim 2 , or any combination thereof.4. The polypeptide of claim 2 , wherein the mutated Loop 1 comprises random substitutions of any combination of alanine claim 2 , tyrosine claim 2 , aspartic acid or serine residues.5. The polypeptide of claim 4 , wherein all Loop 1 residues are replaced by alanine claim 4 , tyrosine claim 4 , aspartic acid or serine residues.6. The polypeptide of claim 4 , wherein the mutated Loop 1 further comprises an extra glycine residue on the C-terminus of Loop 1.7. The polypeptide of claim 2 , wherein the CH2 domain or CH3 domain further comprises a mutated Loop 3 claim 2 , ...

COLLECTION AND METHODS FOR ITS USE

The present disclosure enables methods of identifying the VH and VL class pairs in the human immune repertoire, determining the VH and VL class pairs that are most prevalent and those having favorable biophysical properties. More specifically, the collections of the present disclosure comprise the most prevalent and/or preferred VH and VL class pairings with highly diversified CDRs. 1. A collection of synthetic antibodies or functional fragments thereof , comprising variable heavy chain and variable light chain framework regions , wherein said variable heavy chain framework regions and variable light chain framework regions comprise germline protein sequences of a germline protein pair , wherein said germline protein pair comprises the following properties: i) a relative display rate in Fab format comprising a value within the top 75% of Fabs sampled; ii) an expression level in Fab format of at least 0.4 as compared to Fab VH 1-69 VLA_V11-40 AYA; iii) thermal stability at 600 C or more for at least 45 minutes in Fab format; iv) stability in bovine or mouse serum in Fab format for greater than ten days at 370 C; v) an expression level in IgG format of at least 0.4 as compared to MOR03080; and vi) stability in serum in IgG format for fourteen days at 370 C; wherein said collection of antibodies or functional fragments thereof comprises germline protein sequences of at least two different germline protein pairs , and wherein said germline protein pair is encoded by a germline gene pair.223-. (canceled)24. A collection of nucleic acids encoding the collection according to .25. A vector comprising the nucleic acids according to .26. A recombinant host cell comprising the nucleic acids of .2728-. (canceled)29. A collection of synthetic antibodies or functional fragments thereof claim 24 , comprising variable heavy chain and variable light chain framework regions claim 24 , wherein said framework regions comprise germline protein sequences claim 24 , wherein said germline ...

NOVEL METHODS OF CONSTRUCTING LIBRARIES COMPRISING DISPLAYED AND/OR EXPRESSED MEMBERS OF A DIVERSE FAMILY OF PEPTIDES, POLYPEPTIDES OR PROTEINS AND THE NOVEL LIBRARIES

Methods useful in constructing libraries that collectively display and/or express members of diverse families of peptides, polypeptides or proteins and the libraries produced using those methods. Methods of screening those libraries and the peptides, polypeptides or proteins identified by such screens. 1116.-. (canceled)117. A method for preparing single-stranded nucleic acids , the method comprising the steps of:(i) contacting a single-stranded nucleic acid sequence that has been cleaved with a restriction endonuclease with a partially double-stranded oligonucleotide, the single-stranded region of the oligonucleotide being functionally complementary to the nucleic acids in the region that remains after cleavage, the double-stranded region of the oligonucleotide including any sequences necessary to return the sequences that remain after cleavage into proper and original reading frame for expression and containing a restriction endonuclease recognition site 5′ of those sequences; and(ii) cleaving the partially double-stranded oligonucleotide sequence solely at the restriction endonuclease recognition site contained within the double-stranded region of the partially double-stranded oligonucleotide,the contacting and the cleaving steps being performed at a temperature sufficient to maintain the nucleic acid in substantially single-stranded form, the oligonucleotide being functionally complementary to the nucleic acid over a large enough region to allow the two strands to associate such that cleavage may occur at the chosen temperature and at the desired location, and the cleavage being carried out using a restriction endonuclease that is active at the chosen temperature.118. The method according to claim 117 , wherein the length of the single-stranded portion of the partially double-stranded oligonucleotide is between 2 and 15 bases.119. The method according to claim 118 , wherein the length of the single-stranded portion of the partially double-stranded ...

METHODS OF SCREENING ANTIGEN-BINDING MOLECULES BY NORMALIZING FOR THE CONCENTRATION OF ANTIGEN-BINDING MOLECULE

The invention provides singleplex and multiplex assays for screening of antigen-binding molecules for their affinity to antigens by normalizing for the concentration of the antigen-binding molecule. 1. A method of screening a candidate antigen-binding molecule for affinity to an antigen , comprising:a. contacting the candidate antigen-binding molecule with a substrate, the substrate comprising (i) an antigen immobilized on the substrate, and (ii) a normalization binding molecule immobilized on the substrate;b. detecting a binding of the candidate antigen-binding molecule with the antigen on the substrate;c. detecting a binding of the candidate antigen-binding molecule from (b) to the immobilized normalization binding molecule to determine a concentration of the candidate antigen-binding molecule; andd. determining the affinity of the candidate antigen-binding molecule from the binding of the candidate antigen molecule determined in (b) and the binding of the antigen-binding molecule to the normalization binding molecule determined in (c).2. The method of claim 1 , wherein the candidate antigen-binding molecule is selected from the group consisting of an antibody (Ab) claim 1 , an immunoglobulin G (IgG) claim 1 , an antigen-binding fragment (Fab) claim 1 , variable fragment (Fv) claim 1 , a single-chain fragment variable (scFv) claim 1 , an antibody with one V-gene domain claim 1 , a bivalent diabody claim 1 , and combinations thereof.3. The method of claim 2 , wherein the candidate antigen-binding molecule is an Ab or Fab.4. The method of any of - claim 2 , wherein the candidate antigen-binding molecule is displayed on a bacteriophage (phage).5Escherichia coli, Bacillus pumilus, Bacillus subtilis, Cellulosimicrobium cellulans, Oerskovia turbata, Pseudomonas aeruginosa, Pseudomonas syringae, Salmonella entericaThermus thermophilus.. The method of claim 4 , wherein the phage infects a bacterium selected from the group consisting of and6Escherichia coliE. coli. The ...

HLA-RESTRICTED EPITOPES ENCODED BY SOMATICALLY MUTATED GENES

Mutant epitopes encoded by cancer genes are virtually always located in the interior of cells, making them invisible to conventional antibodies. We generated single chain variable fragments (scFvs) specific for mutant peptides presented on the cell surface by human leukocyte antigen (HLA) molecules. These scFvs can be converted to full-length antibodies, termed MANAbodies, targeting “Mutation Associated Neo-Antigens” bound to HLA. A phage display library representing a highly diverse array of single-chain variable fragment sequences was first designed and constructed. A competitive selection protocol was then used to identify clones specific for peptides bound to pre-defined HLA types. In this way, we obtained scFvs, including one specific for a peptide encoded by a common KRAS mutant and another by a common EGFR mutant. Molecules targeting MANA can be developed that specifically react with mutant peptide-HLA complexes even when these peptides differ by only one amino acid from the normal, wild-type form. 1. An isolated molecule comprising an antibody variable region which specifically binds to a complex of a human leukocyte antigen (HLA) molecule and a peptide which is a portion of a protein ,wherein the peptide comprises a mutant residue, and wherein the mutant residue is in an intracellular epitope of the protein,wherein the molecule does not specifically bind to the HLA molecule when the HLA molecule is not in said complex, andwherein the molecule does not specifically bind to the peptide in its wild-type form.2. The isolated molecule of wherein said complex further comprises a β-2-microglobulin molecule.3. The isolated molecule comprising an antibody variable region of which is an scFv.4. The isolated molecule comprising an antibody variable region of which is a Fab.5. The isolated molecule comprising an antibody variable region of wherein the protein is an oncogenic protein.6. The isolated molecule comprising an antibody variable region of wherein the ...

AMYLOID BETA(1-42) OLIGOMERS, DERIVATIVES THEREOF AND ANTIBODIES THERETO, METHODS OF PREPARATION THEREOF AND USE THEREOF

The invention relates to neuromodulatory oligomers of the amyloid-β(1-42) protein, a particular production method, by means of which the oligomer can be obtained in a reproducible manner at high yield, the use of the oligomers as diagnostic and therapeutics agents, for the generation of oligomer-specific antibodies and for the discovery of substances which can interact with the oligomers and in the formation thereof. Corresponding methods for the production of the antibodies and for discovery of the substances are also disclosed as are the antibodies themselves and the use of the antibodies or substances as diagnostic and therapeutic agents. The invention further relates to derivatives of the oligomers and oligomers based on abbreviated forms of the amyloid-β(1-42) proteins, the production and use thereof. 1. An oligomer of the amyloid β(1-42) protein , having an apparent molecular weight of about 15 kDa in SDS gel electrophoresis , or a derivative thereof.2. An oligomer of the amyloid β(1-42) protein , having an apparent molecular weight of about 20 kDa in SDS gel electrophoresis , or a derivative thereof.3. An oligomer of the amyloid β(1-42) protein , having an apparent molecular weight of about 38 kDa in SDS gel electrophoresis , or a derivative thereof.4. An oligomer of the amyloid β(1-42) protein , having an apparent molecular weight of about 48 kDa in SDS gel electrophoresis , or a derivative thereof.5. A composition , comprising a mixture of an oligomer as claimed in and an oligomer as claimed in , or of derivatives thereof.6. A composition , comprising a mixture of an oligomer as claimed in and an oligomer as claimed in , or of derivatives thereof.7. The oligomer or composition as claimed in claim 1 , characterized in that the derivatives have a detectable label.8. The oligomer or composition as claimed in claim 7 , characterized in that the label is a fluorescent claim 7 , luminescent claim 7 , colorimetric claim 7 , radioactive or magnetic label or is a ...

COMPOSITIONS AND METHODS FOR GROWTH FACTOR MODULATION

Provided herein are proteins, antibodies, assays and methods useful for modulating growth factor levels and/or activities. In some embodiments, such growth factors are members of the TGF-β superfamily of proteins. 116.-. (canceled)17. A method for making a pharmaceutical composition that comprises an antibody , or antigen-binding fragment thereof , that inhibits GDF8 signaling , the method comprising the steps of:i) providing an antigen comprising pro/latent GDF8;ii) subjecting the antigen of step (i) to proteolysis with one or more of furin, bone morphogenetic protein-1 (BMP-1), mammalian tolloid protein (mTLD), mammalian tolloid-like 1 (mTLL1), and mammalian tolloid-like 2 (mTLL2), so as to provide a cleaved antigen;iii) screening for an antibody, or antigen-binding fragment thereof, that specifically binds the cleaved antigen of step (ii), so as to identify a pro/latent GDF8-specific binder;iv) screening for an antibody, or antigen-binding fragment thereof, that is capable of inhibiting GDF8 signaling, so as to identify a GDF8-specific inhibitor; and,v) formulating a recombinant antibody or a variant thereof, or a composition comprising an antigen-binding fragment thereof, identified in steps (iii) and (iv), into a pharmaceutical composition, wherein the recombinant antibody, or antigen-binding fragment thereof, is a fully human or humanized antibody, or antigen-binding fragment thereof.18. The method of claim 17 , wherein the step (iii) comprises screening a library.19. The method of claim 18 , wherein the library is a phage display library.20. The method of claim 17 , further comprising a step of subjecting the recombinant antibody claim 17 , or antigen-binding fragment thereof claim 17 , to affinity maturation.21. The method of claim 17 , wherein the step (iv) comprises screening for an antibody claim 17 , or antigen-binding fragment thereof claim 17 , that inhibits release of mature GDF-8 growth factor from the pro/latent GDF8.22. The method of claim 17 , ...

PHAGE-DISPLAYED SINGLE-CHAIN VARIABLE FRAGMENT LIBRARIES AND USES THEREOF

Disclosed herein is a phage-displayed single-chain variable fragment (scFv) library, which comprises a plurality of phage-displayed scFvs characterized in having a specific CS combination and a specific sequence in each CDR. The present scFv library is useful in efficiently producing different antibodies with binding affinity to different antigens. Accordingly, the present disclosure provides a potential means to generate different antigen-specific antibodies promptly in accordance with the need in experimental researches and/or clinical applications. 1. A phage-displayed single-chain variable fragment (scFv) library comprising a plurality of phage-displayed scFvs , wherein each of the plurality of phage-displayed scFvs comprises a first light chain complementarity determining region (CDR-L1) , a second light chain CDR (CDR-L2) , a third light chain CDR (CDR-L3) , a first heavy chain CDR (CDR-H1) , a second heavy chain CDR (CDR-H2) , and a third heavy chain CDR (CDR-H3) , whereinthe CDR-L1 is encoded by a first coding sequence comprising the nucleic acid sequence of SEQ ID NO: 8 or 10;the CDR-L2 is encoded by a second coding sequence comprising the nucleic acid sequence of SEQ ID NO: 12, 14, 16 or 18;the CDR-L3 is encoded by a third coding sequence comprising the nucleic acid sequence of SEQ ID NO: 20 or 22;the CDR-H1 is encoded by a fourth coding sequence comprising the nucleic acid sequence of SEQ ID NO: 24, 26, 28, 30, 32, 34, 36 or 38;the CDR-H2 is encoded by a fifth coding sequence comprising the nucleic acid sequence of SEQ ID NO: 40 or 42; andthe CDR-H3 is encoded by a sixth coding sequence comprising the nucleic acid sequence of any of SEQ ID NOs: 45-46, 52-56, 60-62 67-70, 76-80, 91-100, 116-130 and 152-172.2. The phage-displayed scFv library of claim 1 , whereinthe first coding sequence has the nucleic acid sequence of SEQ ID NO: 7 or 9;the second coding sequence has the nucleic acid sequence of SEQ ID NO: 11, 13, 15 or 17;the third coding sequence has the ...

Immunoconjugates Comprising Anti-HER2 Antibodies and Pyrrolobenzodiazepines

The invention provides immunoconjugates comprising anti-HER2 antibodies and methods of using the same. 2. The immunoconjugate of claim 1 , wherein the antibody comprises a heavy chain variable region comprising the sequence of SEQ ID NO: 11 and a light chain variable region comprising the sequence of SEQ ID NO: 10.3. (canceled)4. (canceled)5. The immunoconjugate of claim 1 , which is an antibody fragment that binds HER2.6. The immunoconjugate of claim 1 , wherein HER2 is human HER2 comprising amino acids 23 to 1255 of SEQ ID NO: 1.7. The immunoconjugate of an claim 1 , wherein the antibody binds to extracellular domain I of HER2.8. The immunoconjugate of claim 7 , wherein extracellular domain I of HER2 has the sequence of SEQ ID NO: 35.9. The immunoconjugate of claim 4 , which is an IgG1 claim 4 , IgG2a or IgG2b antibody.10. The immunoconjugate of claim 1 , wherein the antibody comprises one or more engineered free cysteine amino acids residues.11. The immunoconjugate of claim 10 , wherein the one or more engineered free cysteine amino acids residues are located in the heavy chain.12. The immunoconjugate of claim 10 , wherein the one or more engineered free cysteine amino acids residues are located in the light chain.13. The immunoconjugate of claim 11 , wherein the antibody comprises at least one mutation in the heavy chain constant region selected from A118C and S400C.14. The immunoconjugate of claim 12 , wherein the antibody comprises at least one mutation in the light chain constant region selected from K149C and V205C.15. The immunoconjugate of claim 1 , wherein the antibody comprises:a) a heavy chain comprising the sequence of SEQ ID NO: 19 and a light chain comprising the sequence of SEQ ID NO: 18; orb) a heavy chain comprising the sequence of SEQ ID NO: 19 and a light chain comprising the sequence of SEQ ID NO: 23; orc) a heavy chain comprising the sequence of SEQ ID NO: 24 and a light chain comprising the sequence of SEQ ID NO: 18.16. The immunoconjugate of ...

HIV Antigens and Antibodies

The present invention relates to a method for reducing the occurrence and/or severity of viral infections. The method embodies procedures for expanding HIV from the blood of HIV antibody negative donors and deriving a non-infectious virus particle product that is antigenic. The procedures for deriving the antigenic, non-infectious virus particle product are optimally designed to maintain the integrity of the envelope proteins while maximizing the depletion of capsid proteins and RNA. The resulting virus particle product, when introduced into humans or non-human animals, enables the production of antibodies that target the natural envelope macromolecular structure that is required for infectivity. The present invention can be applied to producing virus stocks from the blood of HIV-seronegative donors, for deriving non-infectious virus particles that retain intact envelope proteins, for producing anti-viral antibodies, and for administering anti-virus antibodies to patients. 168.-. (canceled)69. A composition comprising polyclonal antibodies , wherein the polyclonal antibodies selectively bind to an epitope on an HIV-1 trimeric envelope glycoprotein subunit (TEGS) of an infectious HIV-1 virus.70. The composition of claim 69 , wherein the TEGS is prepared by a process comprising:obtaining infectious HIV-1 virus particles from human CD4+ cell culture grown in serum-free media;contacting the infectious HIV-1 virus particles with agents that selectively remove from the particle viral RNA and viral capsid protein while retaining viral envelope protein in a non-denatured conformation, wherein the agents do not chemically fix or cross-link the envelope protein; andisolating protein from the contacted infectious HIV-1 virus particles wherein the isolated protein comprises non-infectious complexes comprising a trimeric envelope glycoprotein subunit, the subunit comprising HIV-1 envelope, gp120, and gp41 proteins that are not chemically fixed or cross-linked and substantially ...

ANTIBODY BASED REAGENTS THAT SPECIFICALLY RECOGNIZE TOXIC OLIGOMERIC FORMS OF TAU

The invention relates to antibodies, antibody fragments and binding agents that specifically recognize oligomeric tau but do not bind to monomelic tau, fibrillar tau or non-disease associated forms of tau. 1. An antibody or antibody fragment that specifically recognizes oligomeric tau but does not bind monomeric tau , fibrillar tau or non-disease associated forms of tau.2. The antibody or antibody fragment of claim 1 , wherein the oligomeric tau is dimeric tau or trimeric tau.3. The antibody or antibody fragment of claim 2 , wherein the oligomeric tau is trimeric tau.4. The antibody or antibody fragment of claim 1 , wherein the oligomeric tau is soluble.5. The antibody or antibody fragment of claim 1 , wherein said antibody fragment is isolated according to a method comprising the steps of: (i) a generic protein; and', '(ii) mononeric forms of tau;, 'a. a negative panning of a scFV phage library wherein said negative panning eliminates phage that bind to non-desired antigens wherein said negative panning comprises serially contacting phage withand monitoring the binding of said phage to the generic protein and monomeric forms of tau using Atomic Force Microscope (AFM) Imaging and repeating steps (i) and (ii) until no phage is observed binding to antigen by said AFM imaging to produce an aliquot of phage;b. contacting the aliquot of phage with tau oligomers and incubating for time sufficient to allow binding of phage to said oligomers; andc. eluting the bound phage particles from step (b).6. An antibody or antibody fragment isolated according to a method comprising the steps of: (i) a generic protein; and', '(ii) mononeric forms of tau;', 'and until less than 5% of the phage is observed binding to antigen, which produces an aliquot of phage;, '(a) negative panning a scFV phage library comprising serially contacting phage with(b) positive panning of the aliquot from step (a) comprising contacting the aliquot of phage from step (a) with tau oligomers, and incubating ...

METHOD FOR ISOLATION OF SOLUBLE POLYPEPTIDES

Polypeptides with desirable biophysical properties such as solubility, stability, high expression, monomericity, binding specificity or non-aggregation, including monomeric human Vs and Vs, are identified using a high throughput method for screening polypeptides, comprising the steps of obtaining a phage display library, allowing infection of a bacterial lawn by the library phage, and identifying phage which form larger than average plaques on the bacterial lawn. Sequences of monomeric human Vs and Vs are identified, which may be useful for immunotherapy or as diagnostic agents. Multimer complexes of human Vs and Vs are also identified. The Vs and Vs identified may be used to create further libraries for identifying additional polypeptides. Further, the Vs and Vs may be subjected to DNA shuffling to select for improved biophysical properties. 1. An antigen-binding Vcomprising a FR1 sequence of DIQMTQSPSSLSASVGDRVTITC (amino acids 1-23 of SEQ ID NO:48) , a FR2 sequence of WYQQKPGKAPKLLIF (amino acids 35-49 of SEQ ID NO:48) , a FR3 sequence of GVPSRFSGSGSGTDFTLTISNLQPEDFATYYC (amino acids 57-88 of SEQ ID NO:48) , and a FR4 sequence of FGHGTKVTVL (amino acids 98-107 of SEQ ID NO:48).2. An antigen-binding Vcomprising the FR1 , FR2 , FR3 , and FR4 portion of SEQ ID NO:48 and one or more randomized CDR sequences , wherein one or more of CDR1 , CDR2 and CDR3 of SEQ ID NO:48 is replaced , respectively , with a randomized CDR1 , CDR2 and CDR3.3. The antigen-binding Vof claim 1 , wherein the Vis in a multimeric form.4. The antigen-binding Vof claim 1 , wherein the Vis in a dimeric form.5. The antigen-binding Vof claim 1 , wherein Vis in a trimeric form.6. The antigen-binding Vof claim 1 , wherein the Vis in a pentameric form.7. A display library constructed by preparing nucleic acid sequences coding for the antigen-binding Vof or and expressing the Vso as to display the antigen-binding Vsequence of or .8. The display library of claim 7 , wherein the library is a phage display ...

Antibody variants

Antibody variants of parent antibodies are disclosed which have one or more amino acids inserted in a hypervariable region of the parent antibody and a binding affinity for a target antigen which is at least about two fold stronger than the binding affinity of the parent antibody for the antigen.

PERSONALIZED CANCER IMMUNOTHERAPY

The present disclosure relates to a method for obtaining an antibody or antigen-binding fragment thereof that specifically binds to and against a tumor sample, comprising: administering autologous dendritic cells to a subject; taking immune cells and a tumor sample from the subject; constructing an antibody library of the immune cells; and screening the antibody library to obtain the antibody or a fragment thereof specifically binding to and against the tumor sample. The disclosure also relates to a method for engineering immune cells, an antibody or antigen-binding fragment thereof that specifically binds to and against a tumor sample and uses thereof. 1. A method for obtaining an antibody or antigen-binding fragment thereof that specifically binds to and against a tumor in a subject suffering from the tumor , comprising:taking a tumor sample from the subject;administering dendritic cells to the subject to prime anti-tumor immune response;harvesting immune cells of the subject;constructing an antibody library of the immune cells; andscreening the antibody library to obtain the antibody or a fragment thereof specifically binding to and against the tumor.2. (canceled)3. The method of claim 1 , wherein the subject undergoes surgical resection or simultaneous chemotherapy and radiotherapy before the administration of the dendritic cells.4. The method of claim 1 , wherein the tumor sample is obtained before the administration of the dendritic cells.5. The method of claim 1 , wherein the dendritic cells are autologous.6. (canceled)7. The method of claim 1 , wherein the dendritic cells are administered to the subject more than one time.811.-. (canceled)12. The method of claim 1 , which comprises the following:taking a tumor sample from the subject;administering dendritic cells to the subject twice at 1 week interval to prime anti-tumor immune response after the subject undergoes surgical resection or simultaneous chemotherapy and radiotherapy;harvesting immune cells of ...

Variant nucleic acid libraries for crth2

Provided herein are methods and compositions relating to prostaglandin D2 receptor 2 (DP2 or CRTH2R) libraries having nucleic acids encoding for a scaffold comprising a CRTH2R binding domain. CRTH2R libraries described herein encode for immunoglobulins including antibodies and single domain antibodies. Libraries described herein include variegated libraries comprising nucleic acids each encoding for a predetermined variant of at least one predetermined reference nucleic acid sequence. Further described herein are protein libraries generated when the nucleic acid libraries are translated. Further described herein are cell libraries expressing variegated nucleic acid libraries described herein.

Therapeutic and diagnostic target

The present invention provides methods and compositions for treatment, screening, diagnosis and prognosis of acute myeloid leukemia (AML), B-cell chronic lymphocytic leukemia, breast cancer, colorectal cancer, kidney cancer, head and neck cancer, lung cancer, ovarian cancer or pancreatic cancer, for monitoring the effectiveness of acute myeloid leukemia (AML), B-cell chronic lymphocytic leukemia, breast cancer, colorectal cancer, kidney cancer, head and neck cancer, lung cancer, ovarian cancer or pancreatic cancer treatment, and for drug development. The present invention also provides methods and compositions for depletion of immune cells to treat inflammatory diseases.

PROTEOLYTIC RELEASE OF CELL SURFACE ANTIGENS FOR PHAGE BIOPANNING

The invention described herein features methods of isolating monoclonal antibodies or polypeptides that bind to a cell surface expressed antigen. The method of catch and release utilizes engineered protease site for cleavage antigen-antibody or antigen-polypeptide complexes. In some embodiments the protease cleavage site to cleave the complexes is an exogenous protease to mammalian cell. In various embodiments the protease cleavage site to cleave the complexes is an endogenous protease to mammalian cell. 1. A method of selecting an antibody that binds to a protein of interest , the method comprising the steps of:a) providing a composition comprising a mammalian cell expressing a protein of interest on the cell surface, wherein the protein of interest comprises at least one unique protease cleavage site;b) exposing the mammalian cell to a library of phage-antibody constructs;c) binding one or more phage-antibody constructs from the library to the protein of interest to form one or more antigen-antibody complexes, wherein the antigen is the protein of interest;d) releasing the one or more antigen-antibody complexes from the cell by cleaving the protease cleavage site with a protease;e) propagating the released antigen-antibody complexes and identifying the antibody that binds to the protein of interest;thereby selecting the antibody.2. The method of claim 1 , wherein steps (b) through (e) are repeated.3. The method of claim 2 , wherein steps (b) through (e) are repeated from about 2 to about 10 times.4. The method of claim 1 , wherein unbound phage are removed prior to the repeating of steps (b) through (e).5. The method of claim 1 , wherein the protease is a Tobacco Etch Virus nuclear inclusion endopeptidase (TEVprotease).6. The method of claim 1 , wherein the protease cleavage site is heterologous to the protein of interest.7. The method of claim 1 , wherein the protease cleavage site is endogenous to the protein of interest.8. The method of claim 1 , wherein the ...

SYNTHETIC POLYPEPTIDE LIBRARIES AND METHODS FOR GENERATING NATURALLY DIVERSIFIED POLYPEPTIDE VARIANTS

The invention provides compositions and methods for generating libraries of DNA sequences encoding homologous polypeptides, and uses of the libraries to identify naturally diversified polypeptide variants. The invention also provides compositions and methods for generating collections of synthetic antibody fragments in which one or several complementary determining regions (CDR) are replaced by a collection of the corresponding CDR captured from a natural source. The invention further provides compositions and methods for diversifying a portion of a polypeptide by inserting a diversified sequence of synthetic or natural origin without the need for modification of the original polypeptide coding sequence. 1. A method for producing a collection of nucleic acids , wherein each nucleic acid encodes a human immunoglobulin variable domain comprising a plurality of complementarity determining region 3 (CDR3) sequences isolated separately from the immunoglobulin variable domain repertoire from a mammalian species.2. A method for producing a collection of nucleic acids , wherein each nucleic acid encodes a human immunoglobulin variable domain comprising a plurality of complementarity determining region 3 (CDR3) sequences isolated separately from the immunoglobulin variable domain repertoire from a non-human mammalian species.3. A method for producing a collection of nucleic acids , wherein each nucleic acid encodes a human immunoglobulin variable domain comprising a plurality of complementarity determining region 3 (CDR3) sequences isolated separately from the immunoglobulin variable domain repertoire from a human.4. The method of claim 1 , the method comprising:(a) providing a plurality of Acceptor Framework nucleic acid sequences encoding distinct human immunoglobulin variable domains, each Acceptor Framework nucleic acid sequence comprising a first framework region (FR1), a second framework region (FR2), a third framework region (FR3), and a fourth framework region (FR4), ...

DEVELOPING AN EFFICIENT HYBRIDOMA PLATFORM FOR THERAPEUTIC ANTIBODY DISCOVERY

The instant technology generally relates to improved methods for producing antibodies, antibody libraries, hybridomas, hybridoma libraries, etc. For example, these methods increase the number of antigen-specific B cells produced, increase the number of hybridomas, and/or increase the number of monoclonal antibodies that can be made in a given production cycle. 1. A method for producing an antibody library , the method comprising:(a) injecting one or more animals with an antigen;(b) harvesting draining lymph nodes comprising B cells from each animal;(c) forming a hybridoma between each B cell and a fusion partner; and(d) screening the hybridomas for binding specificity to the antigen; (i) the animals are outbred animals;', '(ii) the animals are injected at multiple sites;', '(iii) the animals are injected every two weeks;', '(iv) the animals are injected for between 6 weeks and 15 weeks;', '(v) multiple adjuvants are used, such that different animals are injected with different adjuvants;', '(vi) B cells are enriched prior to step (c); and/or', '(vii) use of a fusion partner engineered to express both surface and secreted IgG., 'wherein at least two of the following conditions apply2. The method of claim 1 , wherein two of conditions (i)-(vii) apply.3. The method of claim 1 , wherein three of conditions (i)-(vii) apply.4. The method of claim 1 , wherein four of conditions (i)-(vii) apply.5. The method of claim 1 , wherein five of conditions (i)-(vii) apply.6. The method of claim 1 , wherein six of conditions (i)-(vii) apply.7. The method of claim 1 , wherein seven of conditions (i)-(vii) apply.8. The method of claim 1 , wherein at least two of conditions (i)-(vi) apply.9. The method of claim 1 , wherein step (a) comprises injecting two or more animals with an antigen.10. The method of claim 1 , wherein step (a) comprises injecting three or more animals with an antigen.11. The method of claim 1 , wherein step (a) comprises injecting four or more animals with an ...

VARIABLE HEAVY CHAIN ONLY LIBRARIES, METHODS OF PREPARATION THEREOF, AND USES THEREOF

The present disclosure relates to designs and applications of Variable Heavy Only (VHO) domain regions. The present disclosure provides a VHO library of polynucleotides encoding VHO domains, wherein the VHO domains are designed based on the Vh domain of a human Vh family, and a method of generating the VHO library. The present disclosure also provides a phage library displaying the VHO domains encoded by the VHO library through the use of M13 bacteriophage minor coat proteins such as pIX and pVII and a method of generating the phage library. The present disclosure also provides a method of screening the phage library to identify VHO candidates that are capable of binding a target of interest. 1. A VHO library of polynucleotides encoding VHO (variable heavy only) domains , wherein the VHO domains have sequence homology and/or canonical homology with the Vh domain of a human Vh family.2. The VHO library of claim 1 , wherein the human Vh family is IGHV3-23.3. The VHO library of claim 2 , wherein the VHO domains have residue diversity throughout the entire regions of the VHO domains.4. The VHO library of claim 2 , wherein the VHO domains have residue diversity in one or more of the CDR regions.7. The VHO library of claim 5 , wherein position X is replaced with an even distribution of all 20 naturally occurring amino acids except for C for CDR1 claim 5 , C and M for CDR2 claim 5 , and C claim 5 , M claim 5 , and N for CDR3.8. The VHO library of claim 6 , wherein position X is replaced with an even distribution of all 20 naturally occurring amino acids except for C for CDR1 claim 6 , C and M for CDR2 claim 6 , and C claim 6 , M claim 6 , and N for CDR3.9. The VHO library of claim 2 , wherein the VHO domains have residue diversity in one or more of the framework regions.10. The VHO library of claim 2 , wherein one or more of the framework regions have one or more mutations that provide improvements in levels of protein expression claim 2 , protein folding claim 2 , protein ...

VECTORS FOR CLONING AND EXPRESSION OF PROTEINS, METHODS AND APPLICATIONS THEREOF

The present disclosure relates to vectors for cloning and expressing genetic material including but not limiting to antibody gene or parts thereof and methods of generating said vectors. Said vectors express the antibody genes in different formats such as Fab or scFv as a part of intertransfer system, intratransfer system or direct cloning and expression in individual display systems. In particular, phage display technology is used to clone and screen potential antibody genes in phagemid which is followed by the transfer of said genes to yeast vector for further screening and identification of lead molecules against antigens. The present vectors have numerous advantages including uniquely designed inserts/expression cassettes resulting in efficient and smooth transfer of clonal population from phage to yeast vectors resulting in efficient library preparation and identification of lead molecules. 1. A vector construct designed to receive antibody or a fragment thereof from a phagemid comprising at least one cloning region or from a yeast vector comprising at least one cloning region , or , to transfer antibody or a fragment thereof to a yeast vector comprising at least one cloning region , said vector construct containing an expression cassette which comprises:at least one leader sequence;at least one cloning region for receiving a gene encoding a peptide or protein that selectively binds to a biologically active ligand;at least one nucleotide sequence encoding constant region immunoglobulin heavy chain or constant region immunoglobulin light chain, or fragments thereof, or a combination of constant region immunoglobulin heavy chain or fragment thereof and constant region immunoglobulin light chain or fragment thereof, wherein said constant region comprises at least one mutation with respect to constant region of a native immunoglobulin or fragments thereof; andat least one recombinant tag sequence or selection coding nucleic acid sequence;wherein, the at least one ...

SEMI-SYNTHETIC NURSE SHARK VNAR LIBRARIES FOR MAKING AND USING SELECTIVE BINDING COMPOUNDS

The present invention relates to VNAR single chain antibodies and more particularly, to semi-synthetic VNAR libraries derived from nurse shark which may be used to identify individual clones, nucleic acid molecules and polypeptides which encode binding moieties that specifically bind to a cellular target of interest, thereby altering (e.g., antagonizing) target activity in a cell or mimicking the activity of a native molecule. The present invention thus also relates to compounds and compositions comprising a target specific VNAR binding moiety, methods for preparing them, and diagnostic and therapeutic methods of use relating to regulation, e.g., agonism or antagonism of the selected cellular target or target pathway e.g., to treat and/or prevent a pathological condition, disorder or disease in which it is beneficial to alter, e.g., agonize or augment, antagonize, reduce or eliminate the specific cellular target activity. 2. The nucleic acid library of claim 1 , wherein at least 75% of the functional Type 1 VNARs comprise a CDR3 of 26 amino acid residues or more claim 1 , wherein a functional VNAR is non-frameshifted relative to the germline Type 1 VNAR sequence.3. The nucleic acid library of claim 1 , wherein the library comprises from 50 to 2×10or from 50 to 2×10 claim 1 , or more molecules claim 1 , having theoretically distinct nucleic acid sequences.4. The nucleic acid library of claim 1 , wherein said library is a phage display library.5. A method of identifying a polypeptide that binds selectively to a target molecule of interest which comprises:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, '(a) exposing a target molecule of interest to polypeptides produced by expression of a library of ; and'}(b) separating polypeptides that selectively bind from those that do not selectively bind the target molecule.6. The method of claim 5 , wherein the target molecule of interest is expressed on the surface of a phage claim 5 , bacterium or cell claim 5 , or is ...

Collection and Methods For Its Use

The present disclosure enables collections of variable heavy chain and variable light chain pairs comprising, in part, germline protein sequences that are pre-selected for functional properties relevant to developability, wherein the collections may be used to select against any antigen using, for example, phage display.

FIBRONECTIN-BASED BINDING MOLECULES AND USES THEREOF

The invention provides fibronectin type III (Fn3)-based binding molecules that bind to a specific target antigen. The invention further provides bispecific Fn3-based binding molecules that bind to two or more targets simultaneously. The Fn3-based binding molecules of the invention can also be linked together to form multispecific Fn3-based binding molecules, and/or can be conjugated to a non-Fn3 moiety, such as, Human Serum Albumin (HSA), for improved half life and stability. The invention also provides methods for generating, screening and using Fn3-based binding molecules in a variety of therapeutic and diagnostic applications. 143-. (canceled)44. A Fn3-based binding molecule conjugate comprising an Fn3 domain that binds a target and a non-Fn3 moiety which extends half-life of the Fn3-based binding molecule.45. The Fn3-based binding molecule conjugate of claim 44 , wherein the non-Fn3 moiety is selected from the group consisting of human serum albumin (HSA) claim 44 , an antibody Fc region and polyethylene glycol (PEG).46. The Fn3-based binding molecule conjugate of claim 44 , wherein the non-Fn3 moiety is an antibody Fc region.47. The Fn3-based binding molecule conjugate of claim 46 , wherein the antibody Fc region is an Fc region of IgG1.48. The Fn3-based binding molecule conjugate of claim 44 , wherein the Fn3 domain is derived from the wild type tenth Fn3 domain of human fibronectin of SEQ ID NO:1.49. The Fn3-based binding molecule conjugate of claim 48 , wherein the top BC claim 48 , DE and FG loops of the Fn3 domain differ from the wild type Fn3 domain of SEQ ID NO:1.50. The Fn3-based binding molecule conjugate of claim 49 , wherein the top loops differ from the wild type Fn3 domain of SEQ ID NO:1 at one or more of positions 23 claim 49 , 24 claim 49 , 25 claim 49 , 26 claim 49 , 27 claim 49 , 28 claim 49 , 29 claim 49 , 30 claim 49 , 51 claim 49 , 52 claim 49 , 53 claim 49 , 54 claim 49 , 55 claim 49 , 56 claim 49 , 76 claim 49 , 77 claim 49 , 78 claim 49 , ...

METHODS FOR THE GENERATION OF MULTISPECIFIC AND MULTIVALENT ANTIBODIES

The invention provides novel bispecific monoclonal antibodies carrying a different specificity for each binding site of the immunoglobulin molecule and methods for producing novel bispecific monoclonal antibodies carrying a different specificity for each binding site of the immunoglobulin molecule. The antibodies are composed of a single heavy chain and two different light chains, one containing a Kappa constant domain and the other of a Lambda constant domain. The invention provides methods for the isolation of antibodies of different specificities but sharing a common heavy chain. The invention also provides methods for the controlled co-expression of two light chains and a single heavy chain leading to the assembly of monospecific and bispecific antibodies. The invention provides a mean of producing a fully human bispecific and bivalent antibody that is unaltered in sequence and does not involve the use of linkers or other non-human sequences, as well as antibody mixtures of two monospecific antibodies and one bispecific antibody. The invention also provides the means of efficiently purifying the bispecific antibody. 1. A method to generate an antibody mixture comprising three or more monospecific antibodies and three or more bispecific antibodies , all having a common heavy chain , the method comprising:a. Isolating an antibody or antibody fragment region having a specificity determined by a heavy chain variable domain combined with a first light chain variable domain;b. Isolating several antibodies or antibody fragments region having a different specificity determined by the same heavy chain variable domain as the antibody of step a) combined with different light chain variable domains; i. a heavy chain polypeptide comprising the common heavy chain variable domain fused to an immunoglobulin heavy chain constant region;', 'ii. light chain polypeptides comprising all the light chains of the antibodies isolated in step a) and b) fused either to a light chain ...

ANTIBODY LIBRARY AND ANTIBODY SCREENING METHOD USING SAME

The present invention relates to a novel antibody library and an antibody-screening method using same. Having a human sequence-derived specific VH or VL scaffold, the antibody library according to the present invention exhibits high thermodynamic stability and enjoys the advantages of allowing high soluble expression as well as reversible folding. In addition, the antibody according to the present invention includes a variety of rationally controlled CDRs so as to exhibit high specificity and high affinity to all antigens and thus can be advantageously used for selecting an adequate candidate antibody against a target antigen. 1. A set of antibodies or fragments thereof ,wherein each antibody or fragment thereof comprises a pair of a heavy-chain variable region and a light-chain variable region,wherein the heavy-chain variable region comprises:a framework region included in a heavy-chain variable region selected from the group consisting of VH3-15 (SEQ ID NO: 1), VH3-23 (SEQ ID NO: 6) and VH1-69 (SEQ ID NO: 11); anda combination of a heavy-chain complementarity-determining region 1 (CDRH1), a heavy-chain complementarity-determining region 2 (CDRH2), and a heavy-chain complementarity-determining region 3 (CDRH3), which are different for each heavy-chain variable region, andthe light-chain variable region comprises:a framework region included in a light-chain variable region selected from the group consisting of Vκ1-39 (SEQ ID NO: 16), Vκ3-20 (SEQ ID NO: 21), Vκ3-20-2 (SEQ ID NO: 26) and Vλ1-51 (SEQ ID NO: 31); anda combination of a light-chain complementarity-determining region 1 (CDRL1), a light-chain complementarity-determining region 2 (CDRL2), and a light-chain complementarity-determining region 3 (CDRL3), which are different for each light-chain variable region.2. The set of antibodies or fragments thereof according to claim 1 , wherein the set of antibodies or fragments thereof comprises:a framework region included in a pair of heavy- and light-chain variable ...

METHOD OF IMPROVING CHARACTERISTICS OF PROTEINS

The invention provides efficient methods for combining single-substitution libraries of nucleic acids that span and encode proteins of interest and for selecting resultant mutant proteins after expression which have improved properties or characteristics. 18-. (canceled)9. A method of improving a protein binding site , the method comprising the steps of:synthesizing a single substitution library for each of a plurality of domains of a protein binding site, wherein each member of a single substitution library has a nucleotide sequence that encodes amino acid changes at a single amino acid position of its associated domain and that overlaps a nucleotide sequence of at least one member of a different single substitution library, and wherein the domains of the protein binding site are singly substituted at from 1 to 250 amino acid positions;expressing separately each member of each single substitution library as a pre-candidate protein;incubating in a reaction mixture under binding conditions the pre-candidate proteins of each single substitution library with target molecules;washing the target molecules and from pr-candidate proteins remaining bound form a selected library for each domain of the protein;shuffling members of the selected libraries in a PCR to produce a combinatorial shuffled library;expressing members of the shuffled library as candidate proteins;incubating in a reaction mixture under binding conditions the candidate proteins with target molecules;washing the target molecules so that a fraction of candidate proteins remain bound; andselecting members of the shuffled library which encode candidate proteins which remain bound.10. The method of wherein said binding site is that of an antibody or an antibody fragment expressed by a protein display system.11. The method of wherein said protein display system is a yeast display system claim 10 , a mammalian display system claim 10 , a bacterial display system claim 10 , an insect cell display system or a ...

TUMOR SELECTIVE MACROPINOCYTOSIS-DEPENDENT RAPIDLY INTERNALIZING ANTIBODIES

Methods are provided for identifying and selecting antibodies that are internalized into cells via the macropinocytosis pathway. Additionally antibodies that are internalized via this pathway are provided as well as immunoconjugates comprising such antibodies. 1. A method of preparing antibodies that are internalized into a cell by a macropinocytosis pathway , said method comprising:contacting target cells with members of an antibody library and with a marker for macropinocytosis;identifying internalized antibodies that co-localize in said target cells with said marker for macropinocytosis; andselecting those antibodies that co-localize with said marker for macropinocytosis.2. The method of claim 1 , wherein said members of an antibody library are members of a phage display library.3. The method of claim 1 , wherein said members of an antibody library are members of a yeast display library.4. The method according to any one of - claim 1 , wherein said antibody library is an antibody library that is enriched for antibodies that bind to tumor cells.5. The method of claim 4 , wherein said antibody library is an antibody library that is enriched for antibodies that bind to tumor cells and said enrichment is by laser capture microdissection (LCM) of antibodies that bind to tumor cells.6. The method according to any one of - claim 4 , wherein said antibody library is an antibody library that is enriched for antibodies that are internalized into tumor cells.7. The method according to any one of - claim 4 , wherein said marker for macropinocytosis comprises a marker selected from the group consisting of high molecular weight dextran claim 4 , latex beads claim 4 , glass beads claim 4 , Lucifer yellow claim 4 , and soluble enzymes such as horseradish peroxidase.8. The method of claim 7 , wherein said marker for macropinocytosis comprises labeled high molecular weight dextran.9. The method of claim 8 , wherein said marker for macropinocytosis comprises labeled high molecular ...

GPCR BINDING PROTEINS AND SYNTHESIS THEREOF

Provided herein are methods and compositions relating to G protein-coupled receptor (GPCR) libraries having nucleic acids encoding for a scaffold comprising a GPCR binding domain. Libraries described herein include variegated libraries comprising nucleic acids each encoding for a predetermined variant of at least one predetermined reference nucleic acid sequence. Further described herein are protein libraries generated when the nucleic acid libraries are translated. Further described herein are cell libraries expressing variegated nucleic acid libraries described herein. 13. An antibody , wherein the antibody comprises a CDR-H comprising a sequence of any one of SEQ ID NOS: 2420 to 2436.23. The antibody of claim 1 , wherein the antibody comprises a CDR-H comprising a sequence of any one of SEQ ID NOS: 2420 to 2436; and wherein the antibody is a monoclonal antibody claim 1 , a polyclonal antibody claim 1 , a bi-specific antibody claim 1 , a multispecific antibody claim 1 , a grafted antibody claim 1 , a human antibody claim 1 , a humanized antibody claim 1 , a synthetic antibody claim 1 , a chimeric antibody claim 1 , a camelized antibody claim 1 , a single-chain Fvs (scFv) claim 1 , a single chain antibody claim 1 , a Fab fragment claim 1 , a F(ab′)2 fragment claim 1 , a Fd fragment claim 1 , a Fv fragment claim 1 , a single-domain antibody claim 1 , an isolated complementarity determining region (CDR) claim 1 , a diabody claim 1 , a fragment comprised of only a single monomeric variable domain claim 1 , disulfide-linked Fvs (sdFv) claim 1 , an intrabody claim 1 , an anti-idiotypic (anti-Id) antibody claim 1 , or ab antigen-binding fragments thereof.3. A method for treatment of a metabolic disorder claim 1 , comprising administering to a subject in need thereof the antibody of .4. The method of claim 3 , wherein the metabolic disorder is Type II diabetes or obesity.53. A protein library comprising a plurality of proteins claim 3 , wherein each of the proteins of the ...

COMPOSITIONS AND METHODS FOR GROWTH FACTOR MODULATION

Provided herein are proteins, antibodies, assays and methods useful for modulating growth factor levels and/or activities. In some embodiments, such growth factors are members of the TGF-β superfamily of proteins. 1. A method for manufacturing a pharmaceutical composition that comprises an antibody , or an antigen-binding portion thereof , that inhibits TGF-β1 signaling , the method comprising the steps of: a) a homodimer comprising prodomains of transforming growth factor beta (TGF-β1), or prodomain fragments thereof that comprise a straightjacket region that includes at least one cysteine residue present within about the first 4, 5, 6 or 7 N-terminal region; and,', 'b) a protein known to associate with a pro-protein of TGF-β1, selected from the group consisting of: latent TGF-β binding protein 1 (LTBP1), latent TGF-β binding protein 3 (LTBP3), glycoprotein A repetitions predominant (GARP), leucine rich repeat containing 33 (LRRC33), and fragments or variants thereof that comprise two cysteine residues that form disulfide bonds with the straightjacket region of the prodomains, wherein the variants are at least 90% identical to a native sequence;, '(i) providing an antigen comprising a protein complex that comprises(ii) selecting for one or more antibodies, or antigen-binding portions thereof, for the ability to bind the antigen of step (i) and inhibit TGF-β1 signaling; and,(iii) expressing the antibody, or the antigen-binding portion thereof, selected in step (ii) in mammalian cells capable of large scale translation.2. The method of claim 1 , further comprising a step of:(iv) purifying an antibody or antigen-binding portion thereof expressed in step (iii) from the culture medium.3. The method of claim 1 , further comprising a step of:(iv) formulating the antibody or antigen-binding portion thereof purified in step (iv) into a pharmaceutical composition, wherein the antibody, or antigen-binding portion thereof, is a human or humanized antibody or antigen-binding ...

CANCER NEOEPITOPES

Contemplated compositions and methods are directed to cancer neoepitopes and uses of such neoepitopes, especially to generate synthetic antibodies against neoepitopes that may then be employed in the manufacture of a therapeutic agent. Preferred therapeutic agents will comprise a synthetic antibody against a neoepitope, and most preferably in combination with a cellular or non-cellular component for use as a diagnostic or therapeutic agent. 1. A method of generating a pharmaceutical agent for cancer immune therapy , comprising:using matched normal omics data of a tumor to generate in silico a plurality of n-mers that contain at least one patient- and cancer-specific cancer neoepitope;filtering in silico the n-mers to so obtain a subset of neoepitope sequences;preparing at least one synthetic n-mer peptide using sequence information from the subset of neoepitope sequences;using the synthetic n-mer peptide to isolate a recombinant antibody;obtaining sequence information of the complementarity determining region of the recombinant antibody;generating a synthetic antibody using the sequence information of the complementarity determining region of the recombinant antibody; andcoupling the synthetic antibody to a therapeutic or diagnostic agent to so obtain the pharmaceutical agent.2. The method of wherein the matched normal omics data are at least one of whole genomic sequencing data claim 1 , exome sequencing data claim 1 , and transcriptome data.3. The method of wherein the matched normal omics data are matched against normal before treatment of the patient.4. The method of wherein each of the plurality of n-mer peptides has a length of between 7 and 11 amino acids.5. The method of wherein the plurality of n-mer peptides is at least 1 claim 1 ,000 n-mer peptides.6. The method of wherein different of the plurality of n-mer peptides have different neoepitopes.7. The method of wherein the step of filtering includes at least one of filtering by type of mutation claim 1 , ...

ANTI-ROR1 ANTIBODIES

The invention relates to antibodies, and in particular, to antibodies exhibiting specificity for Receptor tyrosine kinase-like Orphan Receptors (ROR), and to uses thereof, for example in the treatment of cancer. The invention extends to polynucleotide and polypeptide sequences encoding the antibodies, and therapeutic uses thereof, and to diagnostic kits comprising these molecules. The invention also extends to antibody-drug conjugates and to uses thereof in therapy. 1. An isolated human anti-Receptor Tyrosine Kinase-Like Orphan Receptor 1 (ROR1) antibody or a functional fragment thereof.2. The isolated antibody or functional fragment thereof according to claim 1 , wherein the functional fragment comprises a fragment selected from a group consisting of VH claim 1 , VL claim 1 , Fd claim 1 , Fv claim 1 , Fab claim 1 , Fab′ claim 1 , scFv claim 1 , F(ab′)and Fc fragment.3. The isolated antibody or functional fragment thereof according to claim 1 , wherein the antibody or functional fragment thereof selectively interacts with ROR1 peptide with an affinity constant of approximately 10to 10M claim 1 , preferably 10to 10M claim 1 , even more preferably claim 1 , 10to 10M.4. The isolated antibody or functional fragment thereof according to claim 1 , wherein the antibody or fragment thereof exhibits an IC50 for ROR1 of about 10to 10M.5. The isolated antibody or functional fragment thereof according to claim 1 , wherein the antibody or fragment thereof is capable of mediating killing of ROR1-expressing tumor cells.6. The isolated antibody or functional fragment thereof according to claim 1 , wherein the antibody or fragment thereof is capable of blocking binding of Wnt5a to ROR1 protein.7. The isolated antibody or functional fragment thereof according to claim 1 , wherein the antibody or fragment thereof is capable of blocking Wnt5a ROR1 phosphorylation.8. The isolated antibody or functional fragment thereof according to claim 1 , wherein the antibody or fragment thereof is ...

Constructs and libraries comprising antibody surrogate light chain sequences

The invention concerns constructs and libraries comprising antibody surrogate light chain sequences. In particular, the invention concerns constructs comprising VpreB sequences, optionally partnered with another polypeptide, such as, for example, antibody heavy chain variable domain sequences, and libraries containing the same.

ANTIGEN-BINDING MOLECULES, THE ANTIGEN-BINDING ACTIVITY OF WHICH VARIES ACCORDING TO THE CONCENTRATION OF COMPOUNDS, AND LIBRARIES OF SAID MOLECULES

An objective of the present invention is to provide target tissue-specific antigen-binding molecules, antigen-binding molecules whose antigen-binding activity varies depending on the concentration of an unnatural compound, libraries comprising a plurality of the antigen-binding molecules which are different from one another, pharmaceutical compositions comprising the antigen-binding molecules, methods of screening for the antigen-binding molecules, and methods for producing the antigen-binding molecules. The present inventors created antigen-binding domains whose antigen-binding activity varies depending on the concentration of a small molecule compound or antigen-binding molecules containing an antigen-binding domain, and libraries comprising a plurality of the antigen-binding domains which are different from one another or antigen-binding domains, and demonstrated that the above-noted objective could be achieved by using the libraries. Various diseases originating from target tissues can be treated in a target tissue-specific manner by using the antigen-binding molecules of the present invention. 119-. (canceled)20. A library that comprises:(i) a plurality of antigen-binding domains having different amino acid sequences from one another, or(ii) a plurality of nucleic acids that encode a plurality of antigen-binding domains having different amino acid sequences from one another, or(iii) a plurality of antigen-binding molecules each comprising an antigen-binding domain, which have different amino acid sequences from one another, or(iv) a plurality of nucleic acids that encode a plurality of antigen-binding molecules each comprising an antigen-binding domain, which have different amino acid sequences from one another,wherein the plurality of antigen-binding domains or antigen-binding domains contained in the plurality of antigen-binding molecules comprises a first antigen-binding domain whose binding affinity for a first antigen varies depending on the concentration ...

NOVEL METHODS OF CONSTRUCTING LIBRARIES COMPRISING DISPLAYED AND/OR EXPRESSED MEMBERS OF A DIVERSE FAMILY OF PEPTIDES, POLYPEPTIDES OR PROTEINS AND THE NOVEL LIBRARIES

Methods useful in constructing libraries that collectively display and/or express members of diverse families of peptides, polypeptides or proteins and the libraries produced using those methods. Methods of screening those libraries and the peptides, polypeptides or proteins identified by such screens. 1116-. (canceled)118. The vector of claim 117 , wherein the DNA sequence set forth in (i) encodes a Fab.119. The vector of claim 117 , wherein the DNA sequence set forth in (i) encodes heavy chain VHCH1.120. The vector of claim 119 , wherein the heavy chain VHCH1 is linked to trpIII.121. The vector of claim 117 , wherein the DNA sequence set forth in (i) encodes light chain VLCL.122. The vector of claim 121 , wherein the light chain VLCL is linked to trpIII.123. The vector of claim 117 , wherein the DNA sequence set forth in (i) encodes scFv.124. The vector of claim 123 , wherein the scFv is VL-VH.125. The vector of claim 123 , wherein the scFv is VH-VL.126. The vector of claim 117 , wherein the DNA sequence encoding the antibody variable region further comprises an inducible promoter.127. The vector of claim 126 , wherein the inducible promoter regulates expression of the DNA sequence encoding the antibody variable region.128. The vector of claim 117 , wherein the DNA sequence encoding the antibody variable region further comprises an amber stop codon.129. The vector of claim 128 , wherein the amber stop codon is located between the antibody variable region and the fragment of the wild-type pIII anchor.130. The vector of claim 117 , wherein the vector is phage or phagemid.131. The vector of claim 117 , wherein the vector further comprises a wild-type gene VIII. This application is a continuation of U.S. patent application Ser. No. 13/464,047, filed May 4, 2012, now U.S. Pat. No. 8,901,045, issued Dec. 2, 2014, which is a continuation of U.S. patent application Ser. No. 10/045,674, filed Oct. 25, 2001, now U.S. Pat. No. 8,288,322, issued Oct. 16, 2012, which is a ...

NOVEL PHAGE DISPLAY LIBRARY, MEMBERS THEREOF AND USES OF THE SAME

The present invention relates to a library (in particular a phage display library) from which an improved human antibody having greater specificity and potency for its target may be generated; and to methods of generating such a human antibody. In particular, the invention relates to a human antibodies against challenging targets obtainable from such a library and which have HCDR3s of at least 18 amino acids in length. 1. A phage library displaying human scFv , wherein at least 50% of the human scFv variable heavy chain CDR3 (CDR H3) displayed by the library is at least 18 amino acids in length.2. The phage library of claim 1 , wherein at least 60% of the human scFv variable heavy chain CDR3 (CDR H3) displayed by the library is at least 18 amino acids in length.3. The phage library of claim 1 , wherein at least 70% of the human scFv variable heavy chain CDR3 (CDR H3) displayed by the library is at least 18 amino acids in length.4. The phage library of claim 1 , wherein at least 80% of the human scFv variable heavy chain CDR3 (CDR H3) displayed by the library is at least 18 amino acids in length.5. The phage library of claim 1 , wherein at least 90% of the human scFv variable heavy chain CDR3 (CDR H3) displayed by the library is at least 18 amino acids in length.6. The phage library of any one of the preceding claims claim 1 , wherein the CDR H3 variable heavy chain is between 18 to 30 amino acids in length.7. The phage library of any one of the preceding claims claim 1 , wherein the CDR H3 variable heavy chain is between 20 to 30 amino acids in length.8. The phage library of any one of the preceding claims claim 1 , wherein the CDR H3 variable heavy chain is between 20 to 24 amino acids in length.9. The phage library of any one of the preceding claims claim 1 , which is generated from a human germline sequence of an antibody having a CDR H3 that is at least 18 amino acids in length.10. The phage library of any one of the preceding claims claim 1 , which is generated ...

Multi-specific monoclonal antibodies

The present invention is relevant to the generation of multi-specific antibodies, antibodies that are distinguished by their ability to bind to multiple antigens with specificity and with affinity. In particular, the present invention is related to bi-specific antibodies.

A METHOD OF GENERATING A SYNTHETIC ANTIBODY LIBRARY, SAID LIBRARY AND APPLICATION(S) THEREOF

The present disclosure relates to a method of generating an antibody library, not limiting to a synthetic antibody gene expression library built on pool of consensus nucleic acid sequences by using codon replacement technology. The present disclosure also relates to a synthetic antibody library generated by employing the method of the present disclosure and application(s) of said antibody library. 2. The synthetic library as claimed in claim 1 , comprising the antibody molecule(s) comprising modified CDR3 of heavy chain (CDRH3) of the antibody molecule(s) having length varying from about 4 amino acids to about 23 amino acids with a specific combination of amino acid frequency claim 1 , wherein:when the length of said CDRH3 is 4 amino acids, frequency of amino acid Glycine at positions 1 and 3 ranges from about 16 to 36%;when the length of said CDRH3 is 5 amino acids, frequency of amino acid Glycine at first 3 positions ranges from about 20 to 38%;when the length of said CDRH3 is 6 amino acids, frequency of amino acid Glycine at first 4 positions ranges from about 11 to 35%;when the length of said CDRH3 is 7 amino acids, frequency of amino acid Glycine at first 4 positions ranges from about 12 to 38%;when the length of said CDRH3 is 8 amino acids, frequency of amino acid Glycine at first 5 positions ranges from about 14 to 34%;when the length of said CDRH3 is 9 amino acids, frequency of amino acid Glycine at first 6 positions ranges from about 12 to 43%;when the length of said CDRH3 is 10 amino acids, frequency of amino acid Glycine at first 7 positions ranges from about 14 to 30%;when the length of said CDRH3 is 11 amino acids, frequency of amino acid Glycine at first 8 positions ranges from about 14 to 28%;when the length of said CDRH3 is 12 amino acids, frequency of amino acid Glycine at first 9 positions ranges from about 15 to 28%;when the length of said CDRH3 is 13 amino acids, frequency of amino acid Glycine at first 10 positions ranges from about 13 to 26%; ...

SYNTHETIC ANTIBODIES TO BAX AND USES THEREOF

Synthetic fragment antigen-binding (Fab) antibodies are disclosed that bind to an N-terminal activation site of BCL-2-associated X-protein (BAX) and inhibit BAX activation. Also disclosed are methods of using the Fabs for measuring inactive monomeric BAX levels, screening for small molecules that bind to an N-terminal activation site of BAX, inhibiting apoptotic cell death, and predicting the ability of a cancer therapy to promote apoptotic cell death. 117-. (canceled)18. A synthetic fragment antigen-binding (Fab) antibody that binds to an N-terminal activation site of BCL-2-associated X-protein (BAX) and inhibits BAX activation ,wherein the synthetic fragment antigen-binding (Fab) antibody is selected from the group consisting of3E8 Fab, having a light chain comprising a CDR L1, a CDRL2, and a CDRL3 region comprising the amino acid sequence QSSYSLI (SEQ ID NO:5), a CDRH1 region comprising the amino sequence LSYYSM (SEQ ID NO:6), a CDRH2 region comprising the amino acid sequence SISPYYGYTY (SEQ ID NO:7), and a CDRH3 region comprising the amino acid sequence RGGAYYFGYYGSGSYAMD (SEQ ID NO:8);3G9 Fab, having a light chain comprising a CDR L1, a CDRL2, and a CDRL3 region comprising the amino acid sequence QHYYYSPWPI (SEQ ID NO:17), a CDRH1 region comprising the amino sequence LYSYYI (SEQ ID NO:18), a CDRH2 region comprising the amino acid sequence SISPYYSSTY (SEQ ID NO:19), and a CDRH3 region comprising the amino acid sequence RSSYSYAGMD (SEQ ID NO:20);2C11 Fab, having a light chain comprising a CDR L1, a CDRL2, and a CDRL3 region comprising the amino acid sequence QSYVSPI (SEQ ID NO:21), a CDRH1 region comprising the amino sequence ISSYYI (SEQ ID NO:22), a CDRH2 region comprising the amino acid sequence SISSYYSSTY (SEQ ID NO:23), and a CDRH3 region comprising the amino acid sequence RVSYGHAYVGYSSGMD (SEQ ID NO:24);3H4 Fab, having a light chain comprising a CDR L1, a CDRL2, and a CDRL3 region comprising the amino acid sequence QSWYYSYPI (SEQ ID NO:25), a CDRH1 region ...

ANTIGEN-SPECIFIC REAGENTS SPECIFIC TO ACTIVE TGF-BETA, METHODS OF PRODUCING THE SAME, AND METHODS OF USE

Antigen-specific reagents (antibody mimetics) that are artificial/mutated proteins and specifically detect active TGF-β. Also provided are methods for producing soluble forms of such antigen-specific reagents (and other affinity reagents), and methods of using the antigen-specific reagents, such as in imaging and flow cytometric analysis. 1. An antigen specific reagent that is a mutated protein and specifically detects active TGF β.2. The antigen-specific reagent of claim 1 , wherein the antigen-specific reagent binds to active TGF β but does not bind to latent TGF-β.3. The antigen-specific reagent of claim 1 , wherein the antigen-specific reagent is soluble.4. The antigen-specific reagent of claim 1 , wherein the antigen-specific reagent is a protein mutated from fibronectin type III domain (FN3) having an amino acid sequence modified at the BC loop as APYGWAPYR (SEQ ID NO: 1) claim 1 , at the DE loop as VPGYYSTA (SEQ ID NO: 2) claim 1 , and at the FG loop as VTGDGPYYQYWFYIS (SEQ ID NO: 3).5. The antigen-specific reagent of claim 1 , wherein the antigen-specific reagent is a protein mutated from fibronectin type III domain (FN3) having an amino acid sequence modified at the BC loop as APAHRYDYYR (SEQ ID NO: 4) claim 1 , at the DE loop as VPPYYGYWYGTA (SEQ ID NO: 5) claim 1 , and at the FG loop as VTHYGGQPYIS (SEQ ID NO: 6).6. The antigen-specific reagent of claim 1 , wherein the antigen-specific reagent is produced by a phage display process and is expressed on an exterior of a membrane.7. The antigen-specific reagent of claim 1 , wherein the antigen-specific reagent comprises a TGF-β binding domain fused with a constant region (Fc) of an immunoglobulin heavy or light chain.8. The antigen-specific reagent of claim 1 , wherein the antigen-specific reagent comprises a TGF-β binding domain fused with rabbit IgG constant region (CH2/CH3).9. A method comprising:producing an antigen-specific reagent that is a mutated protein and specifically detects active TGF β; ...

HUMANIZED ANTI-NUCLEAR ANTIBODIES FOR TARGETING NECROSIS IN CANCER THERAPY

Humanized monoclonal anti-nuclear antibodies with enhanced binding affinity and tumor uptake are presented. Particularly preferred antibodies are site-directed mutants of H-CDR3 with up to about 8-fold improvement in affinity as compared to the non-humanized non-mutated form. In further preferred aspects, such humanized antibodies are employed in tumor necrosis targeted delivery of immune modulators, immune effectors, and other therapeutic or diagnostic agents. 1. A humanized anti-nuclear antibody having an improved binding affinity to a nuclear target relative to the corresponding non-humanized antibody , wherein the humanized antibody has a mutated amino acid in H-CDR3 relative to the corresponding non-humanized antibody , and the H-CDR3 mutated amino acid is selected from the group consisting of Phe→Arg , Ile→His , Gly→Arg , Tyr→Thr , Val→Ser , Arg→Thr , Tyr→Arg , and Arg→Leu.25-. (canceled)6. The humanized antibody of wherein the humanized antibody has at least one ofan H-CDR1 sequence comprising TRYWMH (SEQ ID NO: 1);an H-CDR2 sequence comprising GAIYPGNSDTSYNQKFKG (SEQ ID NO: 2);an H-CDR3 sequence comprising EEIGVRRWFAY (SEQ ID NO: 3);an L-CDR1 sequence comprising RARQSISNYLH (SEQ ID NO: 4);an L-CDR2 sequence comprising ASQSIS (SEQ ID NO: 5); andan L-CDR3 sequence comprising QQSNSWPLT (SEQ ID NO: 6).9. A hybrid molecule comprising at least a binding portion of the antibody of coupled to at least one of a therapeutic agent and an imaging agent.1012-. (canceled)13. A pharmaceutical composition comprising the antibody of or a hybrid molecule comprising at least a binding portion of the antibody coupled to at least one of a therapeutic agent and an imaging agent.14. (canceled)15. A method of targeting a necrotic cell claim 1 , comprising contacting the necrotic cell with an antibody of or the hybrid molecule comprising at least a binding portion of the antibody coupled to at least one of a therapeutic agent and an imaging agent.1617-. (canceled)18. A method of ...

Methods and compositions for ligand directed antibody design

The present disclosure provides, among other things, methods for generating antibodies against a target protein. In some embodiments, a library is provided comprising a plurality of tether antibodies comprising an antigen binding region and a ligand that binds to a target protein. In some embodiments, a library is provided comprising a plurality of candidate antibodies for binding to a target protein.