84 results about "Antibody-Producing Cells" patented technology

Dominant negative alleles of human mismatch repair genes can be used to generate hypermutable cells and organisms. By introducing these genes into cells and transgenic animals, new cell lines and animal varieties with novel and useful properties can be prepared more efficiently than by relying on the natural rate of mutation. These methods are useful for generating genetic diversity within immunoglobulin genes directed against an antigen of interest to produce altered antibodies with enhanced biochemical activity. Moreover, these methods are useful for generating antibody-producing cells with increased level of antibody production. The invention also provides methods for increasing the effector function of monoclonal antibodies and monoclonal antibodies with increased effector function.

A method for identifying and isolating cells which produce secreted proteins. This method is based upon a specific characteristic or the expression level of the secreted protein by transiently capturing the secreted protein on the surface of an individual cell, allowing selection of rare cell clones from a heterogeneous population. Also provided is the use of this method to generate cells which produce a desired level of secreted protein or secreted protein of a particular characteristic(s), and organisms which possess such cells. In particular, the method allows rapid isolation of high expression recombinant antibody-producing cell lines, or may be applied directly to rapid isolation of specific hybridomas, or to the isolation of antibody-producing transgenic animals. This method is applicable for any cell which secretes protein.

A method to produce a cell expressing an antibody from a genomic sequence of the cell comprising a modified immunoglobulin locus using Cre-mediated site-specific recombination is disclosed. The method involves first transfecting an antibody-producing cell with a homology-targeting vector comprising a lox site and a targeting sequence homologous to a first DNA sequence adjacent to the region of the immunoglobulin loci of the genomic sequence which is to be converted to a modified region, so the first lox site is inserted into the genomic sequence via site-specific homologous recombination. Then the cell is transfected with a lox-targeting vector comprising a second lox site suitable for Cre-mediated recombination with the integrated lox site and a modifying sequence to convert the region of the immunoglobulin loci to the modified region. This conversion is performed by interacting the lox sites with Cre in vivo, so that the modifying sequence inserts into the genomic sequence via Cre-mediated site-specific recombination of the lox sites. Also disclosed are a form of the method used to produce a cell expressing a modified antibody molecule using Cre-mediated site-specific recombination, and antibody-producing cells obtainable by the disclosed methods. Class-switching modifications of human antibodies produced in murine hybridoma cells are exemplified.

Dominant negative alleles of human mismatch repair genes can be used to generate hypermutable cells and organisms. Cells may be selected for expression of activation-induced cytidine deaminase (AID), stimulated to produce AID, or manipulated to express AID for further enhancement of hypermutability. These methods are useful for generating genetic diversity within immunoglobulin genes directed against an antigen of interest to produce altered antibodies with enhanced biochemical activity. Moreover, these methods are useful for generating antibody-producing cells with increased level of antibody production.

The present invention relates to compositions and methods for raising antibodies generally comprising 1) providing highly immunogenic vesicles bearing at least one target antigen and 2) immunizing animals with the said antigen-bearing vesicles to induce antigen-specific antibody responses. The invention also relates to methods of screening antibody repertoires comprising 1) providing vesicles bearing at least one target antigen and one marker and 2) isolating antibody-producing cells or particles with defined antigen specificity using the said antigen- and marker-bearing vesicles. Antibodies with defined antigen specificity can then be prepared from isolated antibody-producing cells using known methods of the art. This invention can be used in experimental, research, therapeutic, prophylactic or diagnostic areas.

Provided herein are various processes for the improved production of antibody producing organisms, antibody producing tissues, antibody producing cells and antibodies. In certain embodiments, provided herein are methods for rapidly producing antibody producing organisms, tissues, cells and antibodies derived from humans, organisms, plants or cells that are genetically altered to over-express certain proteins.

This invention generally relates to methods for the production of antibody producing cells and antibodies in protooncogene expressing animals. The invention also relates to methods for the efficient production of antibodies specific for antigens that are normally subject to immunological constraints such as self tolerance. The invention further relates to the production of antibody producing cells and antibodies without the need for the conventional fusing of antibody producing B cells with a myeloma fusion partner.

The invention relates to a method of accelerating the induction of somatic mutations in vitro. The inventive method comprises the expression of at least one cDNA expressing a modified version of the AID gene in the cells to be mutated, in culture conditions and a medium that are suited thereto, said modified version resulting from an AID gene in which the three hydrophobic amino acids, leu189, phe193 and leu196, have been replaced by means of alanine mutations in each case. The invention can be used to induce mutations in Burkitt's lymphoma BL2. The invention can also be used to induce mutations in the immunoglobulin genes of immortalised antibody-producing cells, such as mouse hybridoma cells, human hybridoma cells or human B-cell lines immortalised by the Epstein-Barr virus (EBV).

This invention provides a hybridoma produced by the fusion of a mouse antibody-producing cell and a mouse myeloma which is designated 1-10F-8A and deposited with the ATCC under Accession Number PTA-279, said hybridoma producing a monoclonal antibody which binds to fullerene C60. This invention provides a mouse monoclonal antibody specific for a fullerene-C60 and produced by the mouse monoclonal antibody-producing hybridoma designated 1-10F-8A. The invention provides the amino acid and encoding nucleic acid sequences of the heavy and light chains of the 1-10F-8A monoclonal antibody. This invention also provides methods of determining a serum concentration of a fullerene in a subject and of purifying a fullerene from a sample.

The invention provides methods for fusing a first cell with a second cell to form a hybrid cell. The methods involve incubating a first parental cell producing a first partner of a fusogenic binding partner pair on its surface with a second parental cell producing a second partner of the fusogenic binding partner pair on its surface. In certain embodiments, the parental cells are incubated with a known fusogen such as polyethylene glycol and the fusogenic binding partner pair increases the rate of cell fusion. In many embodiments, the first cell is an antibody producing cell, the second cell is an immortal cell, and the hybrid cell is a hybridoma cell that produces a monoclonal antibody. Also provided by the invention are methods for producing hybridoma cells, and methods for screening those cells for production of a monoclonal antibody of interest. The invention further provides systems and kits for carrying out the subject methods. The subject methods, systems, and kits find use in a variety of different industrial, medical and research applications.

Methods and compounds are disclosed that relate to screening and selection of monoclonal antibodies specific for antigens in heterogeneous antigen mixtures. Antibody-secreting cells such as hybridomas are modified to make them capable of directly binding antigens by capturing their secreted antibody products onto their surface membranes in appropriate binding density and orientation. Selectivity of binding to novel or desired antigens is achieved by first reacting the antigen mixtures affixed to a solid substrate with a polyclonal antibody library that prevents access to the majority of antigens or epitopes other than those that are novel or desired.

The invention relates to a method for preparing an antibody-producing cell line capable of directed constitutive hypermutation of a specific nucleic acid region, comprising the steps of: a) screening a clonal cell population for V gene diversity; b) isolating one or more cells which display V gene diversity and comparing the rate of accumulation of mutations in the V genes and other genes of the selected cells; and c) selecting a cell in which the rate of V gene mutation exceeds that of other gene mutation.

Dominant negative alleles of human mismatch repair genes can be used to generate hypermutable cells and organisms. By introducing these genes into cells and transgenic animals, new cell lines and animal varieties with novel and useful properties can be prepared more efficiently than by relying on the natural rate of mutation. These methods are useful for generating genetic diversity within immunoglobulin genes directed against an antigen of interest to produce altered antibodies with enhanced biochemical activity. Moreover, these methods are useful for generating antibody-producing cells with increased level of antibody production. The invention also provides methods for increasing the affinity of monoclonal antibodies and monoclonal antibodies with increased affinity.

The invention relates to a method for preparing an antibody-producing cell line capable of directed constitutive hypermutation of a specific nucleic acid region, comprising the steps of: a) screening a clonal cell population for V gene diversity; b) isolating one or more cells which display V gene diversity and comparing the rate of accumulation of mutations in the V genes and other genes of the selected cells; and c) selecting a cell in which the rate of V gene mutation exceeds that of other gene mutation.

The invention is a methodology which makes it possible to select from a very large number of cells, a single cell or cells of interest and obtain specific information from those cells in a rapid and efficient manner. As an example of the methodology, a large number of antibody producing cells such as plasma cells are separated so that these individual antibody producing plasma cells are placed in individual wells. The cells are allowed to produce antibodies and the antibodies in the wells are then contacted with a protein bound to a solid surface such as a well top. The protein universally and specifically binds antibodies in the wells. The surface or well tray top includes addresses configured such that each address is specifically related to one of the individual wells containing a cell producing antibodies.

The present invention relates to capsules encapsulating antibody-producing cells, and to use of such capsules and encapsulated cells, respectively, for implantation in vivo for long term delivery or sustained delivery of antibodies of therapeutic interest.

The present invention provides a monoclonal antibody having a pathogenic activity that can induce pemphigus lesion, a peptide specifically recognized by the monoclonal antibody and useful as a therapeutic drug for pemphigus autoimmune disease, etc. As anti-mouse Dsg3 antibody-producing cells are present in the splenocytes of pemphigus vulgaris mouse model constructed by using autoantigen knockout mouse, cell fusion was conducted with the splenocytes of said mouse model and mouse myeloma cells using polyethyleneglycol, hybridomas were constructed, and monoclonal antibodies against Dsg3 were constructed. Among them, a monoclonal antibody having a pathological activity that can induce pemphigus lesions was screened, the base sequence and amino acid sequence in its variable region (heavy chain, light chain) was determined, and the specific epitope part was identified.

The invention provides methods for fusing a first cell with a second cell to form a hybrid cell. The methods involve incubating a first parental cell producing a first partner of a fusogenic binding partner pair on its surface with a second parental cell producing a second partner of the fusogenic binding partner pair on its surface. In certain embodiments, the parental cells are incubated with a known fusogen such as polyethylene glycol and the fusogenic binding partner pair increases the rate of cell fusion. In many embodiments, the first cell is an antibody producing cell, the second cell is an immortal cell, and the hybrid cell is a hybridoma cell that produces a monoclonal antibody. Also provided by the invention are methods for producing hybridoma cells, and methods for screening those cells for production of a monoclonal antibody of interest. The invention further provides systems and kits for carrying out the subject methods. The subject methods, systems, and kits find use in a variety of different industrial, medical and research applications.

The present invention relates to a method for the in vitro production of antibody in an antigen specific manner and, more particularly, to the in vitro production of human antibody in an antigen specific manner. The present invention further extends to the antibody produced therefrom and to the antibody producing cells produced in accordance with the methods disclosed herein. The antibodies and antibody producing cells of the present invention are useful, inter alia, in a range of therapeutic, prophylactic and diagnostic applications.

A method of isolating a monoclonal antibody capable of inhibiting any one of IL-3, GM-CSF and IL-5 binding to the common receptor βc, or a monoclonal antibody capable of inhibiting the cytokines binding to a receptor analogous to βc. The method includes the steps of immunizing an animal with a cytokine receptor or portion of a cytokine containing the critical binding site which portion includes the extracellular domain 4 or analogous domain in the analogous common receptor or part thereof. Antibodies producing cells from the animal are then isolated and fused with a myeloma cell line and then screened for a cell line that produces an antibody of the desired type. A monoclonal antibody, or fragments thereof capable of inhibiting the binding of the cytokines IL-3, GM-CSF and IL-5 to the βc receptor, and a hybridoma cell line producing the antibody are also claimed.

The present invention provides methods for controlling the level of fucosylation and enhancing ADCC activity in antibodies by inhibiting FUT8 expression in antibody-producing cells.

A method for identifying and isolating cells which produce secreted proteins. This method is based upon a specific characteristic or the expression level of the secreted protein by transiently capturing the secreted protein on the surface of an individual cell, allowing selection of rare cell clones from a heterogeneous population. Also provided is the use of this method to generate cells which produce a desired level of secreted protein or secreted protein of a particular characteristic(s), and organisms which possess such cells. In particular, the method allows rapid isolation of high expression recombinant antibody-producing cell lines, or may be applied directly to rapid isolation of specific hybridomas, or to the isolation of antibody-producing transgenic animals. This method is applicable for any cell which secretes protein.

The subject invention relates a method for the production of monoclonal antibodies. The method utilizes an immunized animal having antibody-producing cells with disrupted peripheral tolerance. The invention also provides a method for the use of such monoclonal antibodies, and polyclonal antibodies derived from an immunized animal having antibody-producing cells with disrupted peripheral tolerance, for in vitro and in vivo clinical diagnostics and therapeutics.

The present invention provides a method for producing a fully humanized antibody that recognizes a predetermined antigen and does not rely on a human donor who has been exposed to the antigen. To achieve this, lymphocytes from natural human donors are immunized in vitro with an antigen of interest, and antibody-producing cells directed against said antigen are then identified. Because lymphocytes immunize in vitro rather than in vivo, it is possible to modulate the antigen or antigen fragment recognized by the antibody. A preferred antigen is an HIV gp120 peptide, especially the co-receptor binding region of gp120.

The invention is a methodology which makes it possible to select from a very large number of cells, a single cell or cells of interest and obtain specific information from those cells in a rapid and efficient manner. As an example of the methodology, a large number of antibody producing cells such as plasma cells are separated so that these individual antibody producing plasma cells are placed in individual wells. The cells are allowed to produce antibodies and the antibodies in the wells are then contacted with a protein bound to a solid surface such as a well top. The protein universally and specifically binds antibodies in the wells. The surface or well tray top includes addresses configured such that each address is specifically related to one of the individual wells containing a cell producing antibodies.

The invention discloses a medicinal and edible composition capable of improving the immunity and a preparation method of the medicinal and edible composition. The composition is composed of the medicinal and edible medicinal materials including wheat seedlings, radix ginseng, arillus longan, mulberries, semen sesami nigrum, rhizoma polygonati and the like, as well as fructooligosaccharide and galactooligosaccharide, the integral regulation advantages of the compound traditional Chinese medicine are exerted, the effect of improving the immune functions is obvious, and the experiment proves that the medicinal and edible composition can reduce the toe swelling thickness of normal mice, and improve the activity of the NK cells, the level of the serum hemolysin, the number of antibody-producing cells, the carbon clearance phagocytosis index and the phagocytosis capacity for chicken erythrocytes of macrophagocytes.

Use of a non-human mammalian disease model, wherein the non-human mammal has been implanted with human synovial tissue or other human inflamed tissue containing anti-CCP (cyclic citrullinated peptide) antibody producing cells for (i) analyzing cellular processes in a disease associated with anti-CCP antibodies in such human synovial tissue or other human inflamed tissue, (ii) studying the role of anti-CCP antibodies in the induction and progression of a disease associated with anti-CCP antibodies, (iii) testing the efficacy of a therapeutic agent for the prevention or treatment of a disease associated with anti-CCP antibodies, and (iv) identifying a therapeutic agent useful for the prevention or treatment of a disease associated with anti-CCP antibodies. In one embodiment the non-human mammalian disease model is a mouse, such as a SCID mouse, and the disease associated with anti-CCP antibodies is arthritis, such as RA (rheumatoid arthritis).

Methods are described herein for screening an antibody producing cell within a microfluidic environment. The antibody producing cell may be a B cell lymphocyte, which may be a memory B cell or a plasma cell. An antigen of interest may be brought into proximity with the antibody producing cell and binding of the antigen by an antibody produced by the antibody producing cell may be monitored. Methods of obtaining a sequencing library from an antibody producing cell are also described.