US 11,051,497 B2
Manipulation of immunoglobulin gene diversity and multi-antibody therapeutics
Glenn Friedrich, Cambridge (GB); E-Chiang Lee, Cambridge (GB); Jasper Clube, Cambridge (GB); and Nicholas England, Cambridge (GB)
Assigned to Kymab Limited, Cambridge (GB)
Filed by Kymab Limited, Cambridge (GB)
Filed on Mar. 19, 2014, as Appl. No. 14/220,074.
Application 14/220,074 is a continuation of application No. PCT/GB2012/052298, filed on Sep. 18, 2012.
Claims priority of application No. 1116120 (GB), filed on Sep. 19, 2011; application No. 1116122 (GB), filed on Sep. 19, 2011; application No. 1203257 (GB), filed on Feb. 24, 2012; application No. 1204592 (GB), filed on Mar. 15, 2012; application No. 1205702 (GB), filed on Mar. 29, 2012; application No. 1208749 (GB), filed on May 18, 2012; and application No. 1211692 (GB), filed on Jul. 2, 2012.
Prior Publication US 2014/0212416 A1, Jul. 31, 2014
This patent is subject to a terminal disclaimer.
Int. Cl. A01K 67/027 (2006.01); C07K 16/18 (2006.01); C07K 16/46 (2006.01); C12N 15/85 (2006.01); C07K 16/08 (2006.01); C07K 16/12 (2006.01); C07K 16/10 (2006.01)
CPC A01K 67/0278 (2013.01) [C07K 16/08 (2013.01); C07K 16/12 (2013.01); C07K 16/18 (2013.01); C07K 16/462 (2013.01); C12N 15/8509 (2013.01); A01K 2217/072 (2013.01); A01K 2217/15 (2013.01); A01K 2227/105 (2013.01); A01K 2267/01 (2013.01); C07K 16/085 (2013.01); C07K 16/088 (2013.01); C07K 16/1018 (2013.01); C07K 16/1045 (2013.01); C07K 16/1217 (2013.01); C07K 16/1232 (2013.01); C07K 16/1242 (2013.01); C07K 16/46 (2013.01); C07K 2317/24 (2013.01); C07K 2317/56 (2013.01); C12N 2800/204 (2013.01)] 12 Claims
 
1. A method of isolating an antibody that binds a predetermined antigen, said antibody comprising a human heavy chain variable region, the method comprising
(a) providing a mouse whose genome comprises:
(i) an immunoglobulin heavy (IgH) chain locus comprising a plurality of human VH, one or more human D and one or more human JH gene segments at an endogenous locus upstream of and operably linked to a constant region;
wherein said plurality of human VH gene segments is selected from at least two of the group consisting of: IGHV3-7*01, IGHV3-9*01, IGHV7-4-1*01, IGHV1-3*01, IGHV4-4*02, IGHV3-13*01, IGHV3-23*04 and VH3-20*d01, and where one or more human D gene segments is selected from the group consisting of: IGHD2-2*01, IGHD3-9*01, IGHD3-10*01, IGHD6-13*01, IGHD4-17*01, IGHD6-19*01, IGHD3-22*01 and IGHD1-26*01,
said plurality of human VH gene segments each being capable of joining with a human D gene segment and a human JH gene segment to encode a variable region,
said plurality of human VH gene segments being capable of joining with a human D gene segment and a human JH segment to encode a variable region comprising a HCDR3 of 20 or more amino acids in length in said mouse;
said plurality of human VH gene segments capable of joining with a human D gene segment and a human JH segment to encode a variable region comprising a HCDR3 of 20 or more amino acids in length in said mouse and expresses an IgH heavy chain comprising a HCDR3 of 20 or more amino acids in length;
(ii) an immunoglobulin light (IgL) chain locus comprising one or more human VL gene segments and one or more human JL gene segments upstream of and operatively linked to a constant region;
(b) contacting said mouse with said antigen;
(c) removing B lymphocytes from the mouse and selecting a B lymphocyte expressing antibody that binds to the antigen;
(d) isolating an antibody expressed by the B lymphocytes, wherein in step (d) the antibody which is isolated has an HCDR3 length of at least 20 amino acids.