	US 12,359,001 B2
	Method for producing polypeptide hetero-oligomer
Tomoyuki Igawa, Shizuoka (JP); Hitoshi Katada, Shizuoka (JP); Yuji Hori, Shizuoka (JP); and Shojiro Kadono, Kanagawa (JP)
Assigned to Chugai Seiyaku Kabushiki Kaisha, Tokyo (JP)
Filed by CHUGAI SEIYAKU KABUSHIKI KAISHA, Tokyo (JP)
Filed on Sep. 24, 2021, as Appl. No. 17/483,898.
Application 17/483,898 is a division of application No. 15/562,186, granted, now 11,142,587, previously published as PCT/JP2016/060616, filed on Mar. 31, 2016.
Claims priority of application No. 2015-075448 (JP), filed on Apr. 1, 2015.
Prior Publication US 2022/0010030 A1, Jan. 13, 2022
Int. Cl. C07K 16/46 (2006.01); C07K 16/00 (2006.01); C07K 16/28 (2006.01); G01N 33/15 (2006.01); G01N 33/50 (2006.01); G01N 33/53 (2006.01)

CPC C07K 16/464 (2013.01) [C07K 16/00 (2013.01); C07K 16/2866 (2013.01); C07K 16/468 (2013.01); G01N 33/15 (2013.01); G01N 33/50 (2013.01); G01N 33/53 (2013.01); C07K 2317/31 (2013.01); C07K 2317/526 (2013.01); C07K 2317/53 (2013.01); C07K 2317/55 (2013.01); C07K 2317/56 (2013.01); C07K 2317/565 (2013.01); C07K 2317/624 (2013.01)]

24 Claims

1. A method for producing a heterodimer, the method comprising:

(a) providing a first molecule comprising two copies of a first antibody arm comprising a first antibody heavy chain and a first antibody light chain, wherein the first antibody heavy chain comprises a first heavy chain constant region that comprises a Cys residue and has no core hinge region Cys residues;

(b) providing a second molecule comprising two copies of a second antibody arm comprising a second antibody heavy chain and a second antibody light chain, wherein the second antibody arm is different from the first antibody arm, and wherein the second antibody heavy chain comprises a second heavy chain constant region that comprises a Cys residue and has no core hinge region Cys residues;

(c) incubating the first and second molecules together under a reducing condition that allows Cys-Cys disulfide bonds in the first molecule and in the second molecule to isomerize; and

(d) obtaining a heterodimer comprising the first antibody arm and the second antibody arm, joined by a single Cys-Cys disulfide bond, provided that the heterodimer has no core hinge region disulfide bonds.