US 12,356,967 B2
Immunoglobulin lambda light chain and uses thereof
Andrew J. Murphy, Croton-on-Hudson, NY (US); Lynn Macdonald, Harrison, NY (US); Chunguang Guo, Thornwood, NY (US); and John McWhirter, Hastings-on-Hudson, NY (US)
Assigned to Regeneron Pharmaceuticals, Inc., Tarrytown, NY (US)
Filed by Regeneron Pharmaceuticals, Inc., Tarrytown, NY (US)
Filed on Jun. 1, 2021, as Appl. No. 17/335,727.
Application 17/335,727 is a division of application No. 16/209,820, filed on Dec. 4, 2018, granted, now 11,051,498.
Claims priority of provisional application 62/609,241, filed on Dec. 21, 2017.
Claims priority of provisional application 62/609,251, filed on Dec. 21, 2017.
Claims priority of provisional application 62/594,944, filed on Dec. 5, 2017.
Claims priority of provisional application 62/594,946, filed on Dec. 5, 2017.
Prior Publication US 2021/0292439 A1, Sep. 23, 2021
Int. Cl. A01K 67/0278 (2024.01); A01K 67/0275 (2024.01); C07K 16/00 (2006.01); C07K 16/46 (2006.01); C12N 9/12 (2006.01)
CPC A01K 67/0278 (2013.01) [A01K 67/0275 (2013.01); C07K 16/00 (2013.01); C07K 16/461 (2013.01); C07K 16/462 (2013.01); C12N 9/1264 (2013.01); A01K 2207/15 (2013.01); A01K 2217/052 (2013.01); A01K 2217/072 (2013.01); A01K 2217/15 (2013.01); A01K 2227/105 (2013.01); A01K 2267/01 (2013.01); C07K 2317/10 (2013.01); C07K 2317/14 (2013.01); C07K 2317/21 (2013.01); C07K 2317/24 (2013.01); C07K 2317/515 (2013.01)] 28 Claims
 
1. A method for generating a human λ light chain variable region nucleotide sequence comprising the steps of:
(a) immunizing a genetically modified mouse with an antigen under conditions to produce an immune response to the antigen, wherein the genetically modified mouse has a germline genome comprising:
a first engineered endogenous immunoglobulin κ light chain locus, comprising:
(i) one or more human unrearranged Vλ gene segments,
(ii) one or more human unrearranged Jλ gene segments, and
(iii) a single mouse Cλ gene,
wherein the one or more unrearranged human Vλ gene segments of (i) and the one or more unrearranged human Jλ gene segments of (ii) are in place of the one or more endogenous mouse Vκ gene segments and one or more endogenous Jκ gene segments;
wherein the one or more unrearranged human Vλ gene segments of (i) and the one or more unrearranged human Jλ gene segments of (ii) are operably linked to the single mouse Cλ gene, and
wherein the single mouse Cλ gene of (iii) is operably linked to a mouse kappa enhancer;
wherein the genetically modified mouse lacks a mouse Cκ gene at the first engineered endogenous immunoglobulin κ light chain locus;
wherein the genetically modified mouse comprises a population of B cells that express a population of antibodies specific to the antigen,
wherein the antibodies include immunoglobulin λ light chains that each include a human immunoglobulin light chain variable domain and a mouse Cλ domain,
wherein the mouse Cλ domain of each immunoglobulin λ light chain of the antibodies is expressed from the single mouse Cλ gene; and
(b) isolating one or more B cells from the population of B cells that express the population of antibodies specific to the antigen; and
(c) determining a human λ light chain variable region nucleotide sequence that encodes a human λ light chain variable domain from an antibody from the population of antibodies specific to the antigen generated by the one or more B cells of the genetically modified mouse.