IP Library › Granted Patent US 12,595,316
Granted Patent B2
US 12,595,316 · App. 15/742,803 · Granted Apr 7, 2026

Bispecific and multispecific antibodies and method for isolation of such

Inventors: Richard G. Hibbert (Utrecht, NL); Aran F. Labrijn (Nigtevecht, NL); Arnout F. Gerritsen (Bunnik, NL); Janine Schuurman (Diemen, NL); Paul Parren (Odijk, NL)
Assignee: GENMAB A/S
C07K16/468C07K16/2863C07K16/2887C07K2317/21C07K2317/24C07K2317/31C07K2317/515C07K2317/526C07K2317/732C07K2317/734C07K2317/92C07K2317/94
View Patent ↗
Loading inventors, assignments & file history…
Monitor This Case
Get email alerts when status or documents change.
Order Certified Copies
Most orders are placed with the USPTO same day — all within 24 business hours.
Order via The Patent Place →
Pre-filled with this patent's details
Quick Facts
Patent No.
US 12,595,316
App. No.
15/742,803
Granted
Apr 7, 2026
Kind
B2
Abstract

Novel bispecific and multispecific antibodies having differentially modified light chains, that allows for isolation of the bispecific or multispecific antibodies based on a differential affinity towards an affinity reagent.

Claims (28)

1 . A bispecific or multispecific antibody comprising:

(a) a first light chain/heavy chain pair comprising a first VL region and a first VH region having a first binding specificity and comprising a first kappa light chain constant region and a first heavy chain constant region; and

(b) a second light chain/heavy chain pair comprising a second VL region and a second VH region having a second binding specificity and comprising a second kappa light chain constant region and a second heavy chain constant region;

wherein the first kappa light chain constant region comprises the amino acid substitution V110D or E143D, or both amino acid substitutions V110D and E143D, according to the EU numbering system.

2 . The bispecific or multispecific antibody of claim 1 , wherein the first kappa light chain constant region comprises the amino acid substitution V110D, according to the EU numbering system.

3 . The bispecific or multispecific antibody of claim 1 , wherein the first kappa light chain constant region comprises the amino acid substitution E143D, according to the EU numbering system.

4 . The bispecific or multispecific antibody of claim 1 , wherein the second kappa light chain constant region comprises the amino acid substitution V110D or E143D, or both amino acid substitutions V110D and E143D, according to the EU numbering system.

5 . The bispecific or multispecific antibody of claim 1 , wherein the second kappa light chain constant region comprises the amino acid substitution V110D, according to the EU numbering system.

6 . The bispecific or multispecific antibody of claim 1 , wherein the second kappa light chain constant region comprises the amino acid substitution E143D, according to the EU numbering system.

7 . The bispecific or multispecific antibody of claim 4 , wherein the first and second kappa light chain constant regions do not contain amino acid substitutions in the same positions.

8 . The bispecific or multispecific antibody of claim 1 , which is a bispecific antibody.

9 . The bispecific or multispecific antibody of claim 1 , which is a multispecific antibody.

10 . The bispecific or multispecific antibody of claim 1 , wherein the first kappa light chain constant region comprises both amino acid substitutions V110D and E143D.

11 . The bispecific or multispecific antibody of claim 1 , wherein the second kappa light chain constant region comprises both amino acid substitutions V110D and E143D.

12 . The bispecific or multispecific antibody of claim 1 , wherein the bispecific or multispecific antibody has reduced affinity for an affinity reagent compared to the bispecific or multispecific antibody without the amino acid substitution in said first kappa light chain constant region.

13 . The bispecific or multispecific antibody of claim 2 , which is a bispecific antibody.

14 . The bispecific or multispecific antibody of claim 4 , which is a bispecific antibody.

15 . The bispecific or multispecific antibody of claim 1 , wherein said first and second kappa light chains are human kappa light chains.

16 . A bispecific or multispecific antibody comprising:

(a) a first light chain/heavy chain pair comprising a first VL region and a first VH region having a first binding specificity and comprising a first kappa light chain constant region and a first heavy chain constant region; and

(b) a second light chain/heavy chain pair comprising a second VL region and a second VH region having a second binding specificity and comprising a second kappa light chain constant region and a second heavy chain constant region;

wherein the first kappa light chain constant region comprises an amino acid substitution at a position corresponding to position V110 or position E143, according to the EU numbering system, wherein the amino acid substitution is selected from the group consisting of V110D, V110R, V110E, V110H, V110K, V110N, V110P, V110Q, V110W, and E143D, wherein the second kappa light chain does not comprise an amino acid substitution at a position corresponding to position V110 or position E143, according to the EU numbering system, and wherein the first and second kappa light chain constant regions do not contain amino acid substitutions in the same positions.

17 . The bispecific or multispecific antibody of claim 16 , wherein the first kappa light chain constant region comprises the amino acid substitution V110D, according to the EU numbering system.

18 . The bispecific or multispecific antibody of claim 16 , wherein the first kappa light chain constant region comprises the amino acid substitution E143D, according to the EU numbering system.

19 . The bispecific or multispecific antibody of claim 16 , wherein the first kappa light chain constant region comprises the amino acid substitutions V110D and E143D, according to the EU numbering system.

20 . The bispecific or multispecific antibody of claim 16 , which is a bispecific antibody.

21 . The bispecific or multispecific antibody of claim 16 , which is a multispecific antibody.

22 . The bispecific or multispecific antibody of claim 16 , wherein said first and second kappa light chains are human kappa light chains.

Assignments (3)
SECURITY INTEREST Recorded Dec 15, 2025
From: GENMAB A/S; GENMAB B.V.; GENMAB HOLDING B.V.
To: MORGAN STANLEY SENIOR FUNDING, INC.
Reel/Frame 073933/0597 →
NOTICE OF GRANT OF SECURITY INTEREST IN PATENTS Recorded Dec 15, 2025
From: GENMAB HOLDING B.V.; GENMAB A/S; GENMAB B.V.
To: WILMINGTON TRUST, NATIONAL ASSOCIATION
Reel/Frame 073949/0722 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded May 31, 2018
From: HIBBERT, RICHARD G.; LABRIJN, ARAN FRANK; SCHUURMAN, JANINE; PARREN, PAUL; GERRITSEN, ARNOUT F.
To: GENMAB A/S
Reel/Frame 045954/0246 →
Priority Claims (1)
DK PA 2015 00399 · Jul 9, 2015 · national
Continuity (1)
Related Publication 20180201693A1 · Jul 19, 2018
References Cited (61)
US 5731168A · Carter et al. · 1998 [cited by applicant]
US 8236931B2 · De Wildt · 2012 [cited by examiner]
US 8586713B2 · Davis et al. · 2013 [cited by applicant]
US 20100331527A1 · Davis et al. · 2010 [cited by applicant]
US 20140066599A2 · Blein et al. · 2014 [cited by applicant]
US 20140088295A1 · Smith · 2014 [cited by examiner]
EP 1870459A1 · 2007 [cited by applicant]
EP 2522724A1 · 2012 [cited by applicant]
WO 02100348A2 · 2002 [cited by applicant]
WO 2004035607A2 · 2004 [cited by applicant]
WO 2007110205A2 · 2007 [cited by applicant]
WO 2008052933A2 · 2008 [cited by applicant]
WO 2008119353A1 · 2008 [cited by applicant]
WO 2009080251A1 · 2009 [cited by applicant]
WO 2009089004A1 · 2009 [cited by applicant]
WO 2010151792A1 · 2010 [cited by applicant]
WO 2011131746A2 · 2011 [cited by applicant]
WO 2011147986A1 · 2011 [cited by applicant]
WO 2012143524A2 · 2012 [cited by applicant]
WO 2013060867A2 · 2013 [cited by applicant]
WO WO2013088259A2 · 2013 [cited by examiner]
WO 2013136186A2 · 2013 [cited by applicant]
WO 2014049003A1 · 2014 [cited by applicant]
WO WO2014052717A2 · 2014 [cited by examiner]
Vajdos et al. (J Mol Biol. Jul. 5, 2002;320(2):415-28 (Year: 2002). [cited by examiner]
Brown et al. (J Immunol. May 1996;156(9):3285-91 (Year: 1996). [cited by examiner]
Skolnick et al.(Trends Biotechnol. Jan. 2000;18(1):34-9) (Year: 2000). [cited by examiner]
Burgess et al. (J. Cell Biol. 111:2129-2138) (Year: 1990). [cited by examiner]
Miosge (Proc Natl Acad Sci U S A. Sep. 15, 2015;112(37):E5189-98) (Year: 2015). [cited by examiner]
Bork (Genome Research, 2000, 10:398-400) (Year: 2000). [cited by examiner]
UniProt Accession A2P1G9 (downloaded from https://www.uniprot.org/uniprotkb/A2P1G9/entry on Apr. 6, 2024) (Year: 2024). [cited by examiner]
UniProt PODOX7 (downloaded from https://www.uniprot.org/uniprotkb/PODOX7/entry on Apr. 6, 2024) (Year: 2024). [cited by examiner]
Lindhofer, H. et al., “Preferential species-restricted heavy/light chain pairing in rat/mouse quadromas. Implications for a single-step purification of bispecific antibodies,” J Immunol., vol. 155(1):219-225 (1995). [cited by applicant]
Nilson, B.H.K et al., “Purification of antibodies using protein L-binding framework structures in the light chain variable domain,” Journal of Immunological Methods, vol. 164(1):33-40 (1993). [cited by applicant]
Experimental work: Declaration from WuXi Biologics, 9 pages, Oct. 1, 2024. [cited by applicant]
Fisher, N. et al., “Exploiting light chains for the scalable generation and platform purification of native human bispecific lgG,” Nat Commun., vol. 6 (6113): 12 pages (2014). [cited by applicant]
Graille, M. et al., “Complex between Peptostreptococcus magnus Protein L and a Human Antibody Reveals Structural Convergence in the Interaction Modes of Fab Binding Proteins,” Structure, vol. 9: 679-687 (2001). [cited by applicant]
Gramer, M. et al., “Production of stable bispecific IgG1 by controlled Fab-arm exchange: scalability from bench to large-scale manufacturing by application of standard approaches,” MAbs, vol. 5(6):962-973 (2013). [cited by applicant]
Housden, N.G. et al., “Immunoglobulin-binding domains: Protein L from Peptostreptococcus magnus,” Biochem Soc Trans., vol. 31(Pt 3):716-718 (2003). [cited by applicant]
James, L. et al., “Beta-Edge Interactions in a Pentadecameric Human Antibody Vic, Domain,” J. Mol. Biol., vol. 367:603-608 (2007). [cited by applicant]
Labrijn, A. et al.,“Controlled Fab-arm exchange for the generation of stable bispecific IgG1,” Nat Protoc., vol. 9 (10):2450-2463 (2014). [cited by applicant]
Muzard, J. et al., “Grafting of protein L-binding activity onto recombinant antibody fragments, ”Analytical Biochemistry, vol. 388:331-338 (2009). [cited by applicant]
Rispens, T. et al., “Mechanism of immunoglobulin G4 Fab-arm exchange,” J Am Chem Soc., vol. 133 (26):10302-10311 (2011). [cited by applicant]
Marvin and Zhu, “Recombinant approaches to IgG-like bispecific antibodies,” Acta Pharmacologica Sinica, vol. 26(6):649-658 (2005). [cited by applicant]
Roque, A. et al., “Affinity-based methodologies and ligands for antibody purification: advances and perspectives,” Journal of Chromatography A, vol. 1160:44-45 (2007). [cited by applicant]
Handbook of Therapeutic Antibodies, Second Edition, Chapter 11 Bispecific Antibodies, pp. 267-310 (2014). [cited by applicant]
Experimental work: Summary of products and sequences for Declaration from WuXi Biologics, 2 pages, Oct. 1, 2024. [cited by applicant]
Further Written Submissions, filed in EP Patent No. 3319996, 10 pages, Oct. 20, 2025. [cited by applicant]
Notice of Opposition, filed in EP Patent No. 3319996, 30 pages, Oct. 9, 2024. [cited by applicant]
Response to Notice Notices of Opposition, filed in EP Patent No. 3319996, 58 pages, Feb. 17, 2025. [cited by applicant]
Response to Patentee's Notice of Opposition, filed in EP Patent No. 3319996, 28 pages, Jun. 13, 2025. [cited by applicant]
Response to Proprietor's Submission, filed in EP Patent No. 3319996, 29 pages, May 14, 2025. [cited by applicant]
Second Notice of Opposition, filed in EP Patent No. 3319996, 46 pages, Oct. 9, 2024. [cited by applicant]
Graille, M, et al., “Evidence for Plasticity and Structural Mimicry at the Immunoglobulin Light Chain-Protein L Interface*,” American Society for Biochemistry and Molecular Biology, 277(49), :47500-47506 (2002). [cited by applicant]
Kabat, E. et al., “Sequences of proteins of immunological interest,” US Department of Health and Human Services 5th Edition, 15 pages (1991). [cited by applicant]
Krissinel et al., “Inference of Macromolecular Assemblies from Crystalline State,” J. Mol. Biol., vol. 372: 774-797 (2007). [cited by applicant]
Labrijn, A et al., “Species-specific determinants in the IgG CH3 domain enable Fab-arm exchange by affecting the Noncovalent CH3-CH3 Interaction Strenght, ”J. Immunol., vol. 187(6): 3238-3246 (2011). [cited by applicant]
Labrijn, A.F., et al., “Efficient generation of stable bispecific lgG1 by controlled Fab-arm exchange,” PNAS, vol. 110(13): 5145-5150 (2013). [cited by applicant]
Laemmli U. K., “Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4,” Nature Publishing Group , vol. 227: 680-685 (1970). [cited by applicant]
Lefranc, M-P. et al., “IIMGT, the international ImMunoGeneTics information system 25 years on” Dev Comp Immunol, vol. 27: 55-77 (2003). [cited by applicant]
Nilson, B. et al., “Protein L from Peptostreptococcus magnus binds to the kappa light chain variable domain,” 8 American Society for Biochemistry and Molecular Biology, vol. 267(4): 2234-2239 (1992). [cited by applicant]