US 11,993,642 B2
Trivalent, bispecific antibodies
Ulrich Brinkmann, Weilheim (DE); Rebecca Croasdale, Antdorf (DE); Eike Hoffmann, Seefeld (DE); Christian Klein, Bonstetten (CH); Ekkehard Moessner, Kreuzlingen (CH); Juergen Michael Schanzer, Traunstein (DE); Claudio Sustmann, Munich (DE); and Pablo Umana, Zurich (DE)
Assigned to Hoffmann-La Roche Inc., Little Falls, NJ (US)
Filed by Hoffmann-La Roche Inc., Little Falls, NJ (US)
Filed on May 15, 2019, as Appl. No. 16/413,469.
Application 16/413,469 is a division of application No. 15/857,473, filed on Dec. 28, 2017, abandoned.
Application 15/857,473 is a division of application No. 13/568,224, filed on Aug. 7, 2012, granted, now 9,890,204, issued on Feb. 13, 2018.
Application 13/568,224 is a continuation of application No. 12/752,216, filed on Apr. 1, 2010, abandoned.
Claims priority of application No. 09005108 (EP), filed on Apr. 7, 2009.
Prior Publication US 2020/0062826 A1, Feb. 27, 2020
Int. Cl. C07K 16/00 (2006.01); C07K 16/28 (2006.01); C07K 16/32 (2006.01)
CPC C07K 16/00 (2013.01) [C07K 16/2863 (2013.01); C07K 16/32 (2013.01); C07K 2317/24 (2013.01); C07K 2317/31 (2013.01); C07K 2317/565 (2013.01); C07K 2317/622 (2013.01); C07K 2317/73 (2013.01); C07K 2317/76 (2013.01); C07K 2319/00 (2013.01)] 10 Claims
 
1. A nucleic acid encoding a trivalent, bispecific antibody comprising
a) a full length antibody that specifically binds to a first antigen wherein the full length antibody comprises two antibody heavy chains and two antibody light chains, wherein the CH3 domain of one heavy chain and the CH3 domain of the other heavy chain each meet at an interface which comprises an alteration in the original interface between the antibody CH3 domains, wherein
i) in the CH3 domain of one heavy chain an amino acid residue is replaced with an amino acid residue having a larger side chain volume, thereby generating a protuberance within the interface of the CH3 domain of one heavy chain which is positionable in a cavity within the interface of the CH3 domain of the other heavy chain, and wherein
ii) in the CH3 domain of the other heavy chain an amino acid residue is replaced with an amino acid residue having a smaller side chain volume, thereby generating a cavity within the interface of the second CH3 domain within which the protuberance within the interface of the first CH3 domain is positionable;
b) a polypeptide comprising
ba) an antibody heavy chain variable domain (VH); or
bb) an antibody heavy chain variable domain (VH) and an antibody constant domain 1 (CH1), wherein the N-terminus of the VH domain of the polypeptide is fused via a peptide connector to the C-terminus of the CH3 domain of one of the two heavy chains of the full length antibody;
c) a polypeptide comprising
ca) an antibody light chain variable domain (VL), or
cb) an antibody light chain variable domain (VL) and an antibody light chain constant domain (CL);
wherein the N-terminus of the VL domain of the polypeptide is fused via a peptide connector to the C-terminus of the CH3 domain of the other of the two heavy chains of the full length antibody;
and wherein the antibody heavy chain variable domain (VH) of the polypeptide under b) and the antibody light chain variable domain (VL) of the polypeptide under c) together form an antigen-binding site specifically binding to a second antigen; and
wherein the peptide connectors under b) and c) are peptides with a length between 5 and 50 amino acids.