Method for generating high affinity antibodies
AssigneeRegeneron Pharmaceuticals, Inc.
InventorsGang CHEN, Robert BABB, James P. FANDEL
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Abstract
The present disclosure relates to a method for identifying cells that express antigen-specific antibodies with a high binding affinity for a monomeric antigen. Using fluorescence activated cell sorting, cells expressing high affinity antigen-specific antibodies are selected from a population of immune cells isolated from a mammal that has been immunized with or otherwise exposed to the antigen. Nucleic acids encoding the high affinity antibodies can then be cloned into other lymphoid and non-lymphoid cells where the antibody can be expressed and from which the antibodies can be secreted.
Claims48
-28. (canceled)
A method for selecting primary antibody-producing cells that express antibodies exhibiting high affinity binding for an antigen of interest, the method comprising:
- (a) contacting a population of primary antibody-producing cells that express antibody to the antigen of interest with a monomeric form of the antigen of interest for a time sufficient for the antigen of interest to bind to antibody on the surface of the cells;
- (b) washing the cells for a period of time to remove unbound antigen;
- (c) collecting cells bound to the antigen of interest;
- and
- (d) sorting the cells collected in step (c) to obtain cells that express antibodies of high affinity, wherein the high affinity antibodies have a dissociation equilibrium constant (KD) of 0.1 pM to 25 nM.
Dependencies are inferred from the claim text.