US 11,808,766 B2
Method for generating high affinity antibodies
Gang Chen, Yorktown Heights, NY (US); Robert Babb, River Edge, NJ (US); and James P. Fandl, LaGrangeville, NY (US)
Assigned to Regeneron Pharmaceuticals, Inc., Tarrytown, NY (US)
Filed by Regeneron Pharmaceuticals, Inc., Tarrytown, NY (US)
Filed on Aug. 7, 2020, as Appl. No. 16/987,639.
Application 16/987,639 is a continuation of application No. 15/526,522, granted, now 10,752,698, previously published as PCT/US2015/060523, filed on Nov. 13, 2015.
Claims priority of provisional application 62/079,899, filed on Nov. 14, 2014.
Prior Publication US 2020/0369783 A1, Nov. 26, 2020
This patent is subject to a terminal disclaimer.
Int. Cl. C07K 16/00 (2006.01); G01N 33/569 (2006.01); C07K 16/22 (2006.01); C07K 16/32 (2006.01); G01N 33/50 (2006.01)
CPC G01N 33/56972 (2013.01) [C07K 16/00 (2013.01); C07K 16/22 (2013.01); C07K 16/32 (2013.01); G01N 33/50 (2013.01); C07K 2317/10 (2013.01); C07K 2317/92 (2013.01); C12N 2510/02 (2013.01)] 47 Claims
 
1. 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, wherein the concentration of the monomeric antigen of interest is between 0.1-2 nM or between 5-10 nM;
(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.