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Publication Number
US-12595592-B2
Publication Date
2026-04-07
Expiration Date
Abstract
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.
Core Innovation
The invention describes a method of generating and identifying a recombinant antibody that binds at least one target antigen by using cDNA amplified from mRNA of B cells from a human or a non-human animal selected from rabbit, mouse, rat, hamster, guinea pig, and goat. The cDNA library prepared in this way comprises cDNA encoding V_H and V_L domains, and supports preparation of a recombinant immunoglobulin library based on V_H and V_L domains derived from antigen-capable B cells.
The method generates antibodies from the V_H and V_L domains in a eukaryotic cell production host, where the antibodies comprise light chain/heavy chain combinations. After generation, antibodies are screened with the at least one target antigen to identify a subset of antibodies capable of binding to the at least one target antigen.
The method further expresses at least one of the identified subset of antibodies in the same eukaryotic cell production host as was used in the generating step. This same-host expression is used whereby the recombinant antibody that binds at least one target antigen is identified and generated, and the framework supports comprehensive monoclonal antibody generation without hybridomas.
Claims Coverage
The independent claim is clm-00001. It covers four inventive features in a connected sequence: preparing a V_H and V_L cDNA library from B-cell mRNA, generating light chain/heavy chain antibody combinations in a eukaryotic cell production host, screening with the target antigen to select binders, and expressing identified subset antibodies in the same host to identify and generate recombinant antibodies that bind at least one target antigen.
Amplifying cDNA from B cells to prepare a V_H and V_L cDNA library
cDNA obtained from mRNA of B cells from a human or a non-human animal selected from rabbit, mouse, rat, hamster, guinea pig, and goat is amplified to prepare a cDNA library comprising cDNA encoding V_H and V_L domains.
Generating light chain/heavy chain combinations in a eukaryotic cell production host
Antibodies are generated from the V_H and V_L domains in a eukaryotic cell production host, whereby the antibodies comprise light chain/heavy chain combinations.
Screening antibodies with the target antigen to identify a binding subset
The antibodies are screened with the at least one target antigen to identify a subset of antibodies capable of binding to the at least one target antigen.
Expressing identified subset antibodies in the same eukaryotic cell production host
At least one antibody of the identified subset is expressed in the same eukaryotic cell production host as was used in the generating step, whereby the recombinant antibody that binds at least one target antigen is identified and generated.
Independent claim clm-00001 provides a connected sequence of steps that ties together V_H/V_L cDNA library preparation from B-cell mRNA, antibody generation in a eukaryotic cell production host, target-antigen screening to select binding subsets, and same-host expression to identify and generate recombinant antibodies binding at least one target antigen.
Stated Advantages
Documented Applications
No documented applications found
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