Method of enhancing of binding activity of antibody composition to Fcγ receptor IIIa

• US 8,313,913 B2
• Filed: 08/28/2009
• Issued: 11/20/2012
• Est. Priority Date: 04/09/2002
• Status: Active Grant

• Current Assignee: Kyowa Kirin Co., Ltd. (Kirin Holdings Company Limited)
• Original Assignee: Kyowa Hakko Kirin Company Limited (Kirin Holdings Company Limited)
• Inventors: Nakamura, Kazuyasu, Shitara, Kenya
• Primary Examiner(s): Kelly, Robert M
• Application Number:
  US12/550,160
• Publication Number:
  US 20100092997A1
• Time in Patent Office: 1,180 Days
• Field of Search: None
• US Class Current: 435/7.1
• CPC Class Codes: A61K 39/395 Antibodies agglutinins A61K38/36 ; as drug carriers A61K47/50; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 31/14 for RNA viruses
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: A61P 37/08 Antiallergic agents antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C07K 16/22 against growth factors ; against growth regulators
• CPC Class Codes: C07K 16/283 against Fc-receptors, e.g. CD16, CD32, CD64 CD23 C07K16/2851
• CPC Class Codes: C07K 16/2866 against receptors for cytokines, lymphokines, interferons
• CPC Class Codes: C07K 16/3084 against tumour-associated gangliosides
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/41 Glycosylation, sialylation, or fucosylation
• CPC Class Codes: C07K 2317/732 Antibody-dependent cellular cytotoxicity [ADCC]
• CPC Class Codes: C07K 2319/30 Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
• CPC Class Codes: C12P 21/02 having a known sequence of two or more amino acids, e.g. glutathione

Abstract

A method for enhancing a binding activity of an antibody composition to Fcγ receptor IIIa, which comprises modifying a complex N-glycoside-linked sugar chain which is bound to the Fc region of an antibody molecule; a method for enhancing an antibody-dependent cell-mediated cytotoxic activity of an antibody composition; a process for producing an antibody composition having an enhanced binding activity to Fcγ receptor IIIa; a method for detecting the ratio of a sugar chain in which fucose is not bound to N-acetylglucosamine in the reducing end in the sugar chain among total complex N-glycoside-linked sugar chains bound to the Fc region in an antibody composition; an Fc fusion protein composition produced by using a cell resistant to a lectin which recognizes a sugar chain in which 1-position of fucose is bound to 6-position of N-acetylglucosamine in the reducing end through α-bond in a complex N-glycoside-linked sugar chain; and a process for producing the same.

Claims

1. A method for detecting the antibody-dependent cell-mediated cytotoxic (ADCC) activity of an antibody composition, said antibody molecules having a sugar chain, wherein at least part of the antibody molecule has a no-fucose sugar chain in which fucose is not bound to N-acetylglucosamine in the reducing end of the sugar chain of the antibody molecule, the method comprising:

(a) preparing a standard curve from the binding activities of three or more reference antibody compositions to a Fcγ

receptor IIIa, wherein the no-fucose sugar chain ratio and the ADCC activity of each reference antibody composition is known, comprising (i) contacting each reference antibody composition, separately, with an antigen which binds to the variable regions of the antibodies in each reference antibody composition, to form a reference antibody-antigen complex for each reference antibody composition;

(ii) contacting each reference antibody-antigen complex with Fcγ

receptor IIIa; and

(iii) measuring the binding activity of each reference antibody-antigen complex to the Fcγ

receptor IIIa;

(b) determining the binding activity of a test antibody composition, comprising (i) contacting a test antibody composition with said antigen, wherein said antigen bindgs to the variable regions of the antibodies in said test antibody composition, to form a test antibody-antigen complex;

(ii) contacting the test antibody-antigen complex to the Fcγ

receptor IIIa; and

(iii) measuring the binding activity of the test antibody-antigen complex to the Fcγ

receptor IIIa; and

(c) determining an ADCC activity for the test antibody composition by comparing the binding activity of the test antibody-antigen complex to the Fcγ

receptor IIIa with a standard curve of the binding activities of the reference antibody-antigen complexes to the Fcγ

receptor IIIa.