US 10752698

Patent Overview: US10752698B2 — "Method for Generating High Affinity Antibodies"

Source: https://patents.google.com/patent/[US10752698](/patent/US10752698)/en

Bibliographic data (as supplied): Patent number US10752698B2; application US15/526,522; filed 2015-11-13; priority date 2014-11-14; granted/publication date 2020-08-25; current assignee Regeneron Pharmaceuticals Inc (original assignee same); inventors Gang Chen, Robert Babb, James P. Fandl. Listed legal status: Active, adjusted expiration 2036-03-24 (the source notes legal status is an assumption, not a legal conclusion).

Purpose: High affinity is described as desirable for therapeutic antibodies, but very high affinity antibodies are rare and standard methods (e.g., mouse hybridomas, display libraries) are inefficient at isolating them. The invention is based on the observation that cells expressing high-affinity antibody can be selected and enriched directly from a population of antibody-producing cells of varying affinities, prior to single-cell isolation.

Plain-language scope: The method involves contacting a population of primary antibody-producing cells with a low concentration of labeled antigen; washing the labeled antigen-bound cells with buffer for about 15–60 minutes; then isolating the antigen-bound cells. Isolation may use a labeled antigen-binding protein and fluorescence-activated cell sorting (FACS) to identify single cells expressing high-affinity antibody. Labeled antigen concentrations are described as about 1–15 nM, about 2–10 nM, or about 5 nM; biotinylated antigen with fluorescently labeled streptavidin is described. The method is said to separate high-affinity from low-affinity antibodies (e.g., K_D less than about 25, 20, 15, or 10 nM) and to work with plasma cells, B cells, hybridomas, other recombinant cells, and tissue-derived cells. Host cells comprising nucleic acid encoding isolated antibodies are also contemplated. Figures 1–2 show bead and hybridoma separations by flow cytometry.

What the excerpt does not establish: It does not provide complete claim language (only "prior art keywords" and summary passages are supplied), the full specification, examples beyond partial figure descriptions, prosecution history, or any litigation, ownership-transfer, validity, or prior-art analysis. Any statement requiring those materials is not supported here.