# US 12,595,316 B2 · Bispecific and multispecific antibodies and method for isolation of such > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 12,595,316 B2 - Document ID: US12595316B2 - Status: Granted - Kind: B2 - Application number: 15/742,803 - Filed: 2016-07-01 - Published: 2018-07-19 - Granted: 2026-04-07 - Earliest claimed: 2015-07-09 - Assignees: Genmab A/S - Applicants: Genmab A/S - Inventors: Richard G. Hibbert; Aran F. Labrijn; Arnout F. Gerritsen; Janine Schuurman; Paul Parren - Web page: https://patentgrep.com/patent/US12595316B2 ## CPC - C07K 16/468: {Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies} - C07K 16/2863: {against receptors for growth factors, growth regulators} - C07K 16/2887: {against CD20} - C07K 2317/21: from primates, e.g. man - C07K 2317/24: containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered - C07K 2317/31: multispecific - C07K 2317/515: Complete light chain, i.e. VL + CL - C07K 2317/526: CH3 domain - C07K 2317/732: Antibody-dependent cellular cytotoxicity [ADCC] - C07K 2317/734: Complement-dependent cytotoxicity [CDC] - C07K 2317/92: Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value - C07K 2317/94: Stability, e.g. half-life, pH, temperature or enzyme-resistance - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) ## 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 ### Claim 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. ### Claim 2 (depends on 1) 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. ### Claim 3 (depends on 1) 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. ### Claim 4 (depends on 1) 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. ### Claim 5 (depends on 1) 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. ### Claim 6 (depends on 1) 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. ### Claim 7 (depends on 4) 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. ### Claim 8 (depends on 1) The bispecific or multispecific antibody of claim 1, which is a bispecific antibody. ### Claim 9 (depends on 1) The bispecific or multispecific antibody of claim 1, which is a multispecific antibody. ### Claim 10 (depends on 1) The bispecific or multispecific antibody of claim 1, wherein the first kappa light chain constant region comprises both amino acid substitutions V110D and E143D. ### Claim 11 (depends on 1) The bispecific or multispecific antibody of claim 1, wherein the second kappa light chain constant region comprises both amino acid substitutions V110D and E143D. ### Claim 12 (depends on 1) 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. ### Claim 13 (depends on 2) The bispecific or multispecific antibody of claim 2, which is a bispecific antibody. ### Claim 14 (depends on 4) The bispecific or multispecific antibody of claim 4, which is a bispecific antibody. ### Claim 15 (depends on 1) The bispecific or multispecific antibody of claim 1, wherein said first and second kappa light chains are human kappa light chains. ### Claim 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. ### Claim 17 (depends on 16) 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. ### Claim 18 (depends on 16) 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. ### Claim 19 (depends on 16) 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. ### Claim 20 (depends on 16) The bispecific or multispecific antibody of claim 16, which is a bispecific antibody. ### Claim 21 (depends on 16) The bispecific or multispecific antibody of claim 16, which is a multispecific antibody. ### Claim 22 (depends on 16) The bispecific or multispecific antibody of claim 16, wherein said first and second kappa light chains are human kappa light chains.