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

• US 20040259150A1
• Filed: 04/09/2003
• Published: 12/23/2004
• Est. Priority Date: 04/09/2002
• Status: Abandoned Application

• Current Assignee: Kyowa Hakko Kirin Company Limited (Kirin Holdings Company Limited)
• Original Assignee: Kyowa Hakko Kogyo Company Limited (Kirin Holdings Company Limited)
• Inventors: Nakamura, Kazuyasu, Shitara, Kenya
• Application Number:
  US10/409,611
• Publication Number:
  US 20040259150A1
• Time in Patent Office: Days
• 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 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.

2. The method according to claim 1, wherein the modification of a complex N-glycoside-linked sugar chain which is bound to the Fc region of an antibody molecule is to bind a sugar chain in which 1-position of fucose is not bound to 6-position of N-acetylglucosamine in the reducing end through α

-bond in the complex N-glycoside-linked sugar chain to the Fc region in the antibody molecule.

3. The method according to claim 1, wherein the sugar chain is synthesized by a cell in which the activity of a protein relating to modification of a sugar chain in which fucose is bound to N-acetylglucosamine in the reducing end in the complex N-glycoside-linked sugar chain is decreased or deleted.

4. The method according to claim 3, wherein the protein relating to

(a) a protein relating to synthesis of an intracellular sugar nucleotide, GDP-fucose;

(b) a protein relating to modification of 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;

(c) a protein relating to transport of an intracellular sugar nucleotide, GDP-fucose to the Golgi body.

5. The method according to claim 3, wherein the cell is resistant to a lectin which recognizes a sugar chain structure 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.

6. The method according to claim 3, wherein the cell is resistant to at least one lectin selected from the group consisting of the following (a) to (d):

(a) a Lens culinaris lectin;

7. The method according to claim 3, wherein the cell is selected from the group consisting of a yeast, an animal cell, an insect cell and a plant cell.

8. The method according to claim 3, wherein the cell is selected from the group consisting of the following (a) to (i):

(a) a CHO cell derived from a Chinese hamster ovary tissue;

(b) a rat myeloma cell line YB2/3HL.P2.G11.16Ag.20 cell;

(c) a BHK cell derived from a Syrian hamster kidney tissue;

(d) a mouse myeloma cell line NS0 cell;

(e) a mouse myeloma cell line SP2/0-Ag14 cell;

(f) a hybridoma cell;

(g) a human leukemic cell line Namalwa cell;

(h) an embryonic stem cell;

(i) a fertilized egg cell.

9. The method according to claim 1,

(c) an antibody fragment comprising the Fc region of (a) or (b);

(d) a fusion protein comprising the Fc region of (a) or (b).

10. The method according to claim 1, wherein the antibody molecule belongs to an IgG class.

11. The method according to claim 1, wherein, in the complex N-glycoside-linked sugar chain which is bound to the Fc region of an antibody molecule, the ratio of a sugar chain in which 1-position of fucose is not bound to 6-position of N-acetylglucosamine in the reducing end through α

-bond is 20% or more of total complex N-glycoside-linked sugar chains.

12. A method for enhancing an antibody-dependent cell-mediated cytotoxic activity of an antibody composition, which comprises using the method according to claim 1.

24. A process for producing an antibody composition having an increased antibody-dependent cell-mediated cytotoxic activity, which comprises using the process according to claim 12.

13. A process for producing an antibody composition having an

14. The process according to claim 13, wherein the modification of a complex N-glycoside-linked sugar chain which is bound to the Fc region of an antibody molecule is to bin a sugar chain in which 1-position of fucose is not bound to 6-position of N-acetylglucosamine in the reducing end through α

-bond in the complex N-glycoside-linked sugar chain to the Fc region in the antibody molecule.

15. The process according to claim 13, wherein the sugar chain is synthesized by a cell in which the activity of a protein relating to modification of a sugar chain in which fucose is bound to N-acetylglucosamine in the reducing end in the complex N-glycoside-linked sugar chain is decreased or deleted.

16. The process according to claim 15, wherein the protein relating to modification of a sugar chain in which fucose is bound to N-acetylglucosamine in the reducing end in the complex N-glycoside-linked sugar chain is selected from the group consisting of the following (a), (b) and (c):

(a) a protein relating to synthesis of an intracellular sugar nucleotide, GDP-fucose;

(c) a protein relating to transport of an intracellular sugar nucleotide, GDP-fucose to the Golgi body.

17. The process according to claim 15, wherein the cell is resistant to a lectin which recognizes a sugar chain structure 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.

18. The process according to claim 15, wherein the cell is resistant to at least one lectin selected from the group consisting of the following (a) to (d):

(a) a Lens culinaris lectin;

(b) a Pisum sativum lectin;

(c) a Vicia faba lectin;

(d) an Aleuria aurantia lectin.

19. The process according to claim 15

20. The process according to claim 15, wherein the cell is selected from the group consisting of the following (a) to (i):

(a) a CHO cell derived from a Chinese hamster ovary tissue;

(b) a rat myeloma cell line YB2/3HL.P2.G11.16Ag.20 cell;

(c) a BHK cell derived from a Syrian hamster kidney tissue;

(d) a mouse myeloma cell line NS0 cell;

(e) a mouse myeloma cell line SP2/0-Ag14 cell;

(f) a hybridoma cell;

(g) a human leukemic cell line Namalwa cell;

(h) an embryonic stem cell;

(i) a fertilized egg cell.

21. The process according to claim 13, wherein the antibody molecule is selected from the group consisting of the following (a) to (d):

(a) a human antibody;

(b) a humanized antibody;

22. The process according to claim 13, wherein the antibody molecule belongs to an IgG class.

23. The method according to claim 13, wherein, in the complex N-glycoside-linked sugar chain which is bound to the Fc region of an antibody molecule, the ratio of a sugar chain in which 1-position of fucose is not bound to 6-position of N-acetylglucosamine in the reducing end through α

-bond is 20% or more of total complex N-glycoside-linked sugar chains.

25. An antibody composition produced by the process according to claim 13.

26. A method for detecting the ratio of a sugar chain in which with an Fcγ

receptor IIIa to measure the binding activity to the Fcγ

receptor IIIa; and

detecting the ratio of a sugar chain in a standard antibody composition with a standard curve showing the binding activity to the Fcγ

receptor IIIa.

27. A method for detecting the antibody-dependent cell-mediated cytotoxic activity in an antibody composition, which comprises:

reacting an antigen with a tested antibody composition to form a complex of the antigen and the antibody composition;

contacting the complex with an Fcγ

receptor IIIa to measure the binding activity to the Fcγ

receptor IIIa; and

detecting the ratio of a sugar chain in a standard antibody composition with a standard curve showing the binding activity to the Fcγ

receptor IIIa.

28. 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, which comprises:

contacting a tested antibody composition with a Fcγ

receptor IIIa to measure the binding activity of the antibody composition to the Fcγ

29. A method for detecting the antibody-dependent cell-mediated cytotoxic activity in an antibody composition, which comprises:

contacting a tested antibody composition with a Fcγ

receptor IIIa to measure the binding activity of the antibody composition to the Fcγ

receptor IIIa; and

detecting the ratio of a sugar chain in a standard antibody composition with a standard curve showing the binding activity to the Fcγ

receptor IIIa.

30. An Fc fusion protein composition produced by using a cell resistant to a lectin which recognizes a sugar chain structure 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.

31. The Fc fusion protein composition according to claim 30, wherein the cell is selected from the group consisting of the following (a), (b) and (c):

(a) an enzyme protein relating to synthesis of an intracellular sugar nucleotide, GDP-fucose;

(b) an enzyme protein relating to modification of a sugar chain in which 1-

wherein the activity of the protein is decreased or deleted.

32. The Fc fusion protein composition according to claim 30, wherein the cell is resistant to at least one lectin selected from the group consisting of the following (a) to (d):

(a) a Lens culinaris lectin;

(b) a Pisum sativum lectin;

(c) a Vicia faba lectin;

(d) an Aleuria aurantia lectin.

33. The Fc fusion protein composition according to claim 30, wherein the cell is a cell into which a gene encoding an Fc fusion protein is introduced.

34. The Fc fusion protein composition according to claim 33, wherein the Fc is derived from an IgG class of an antibody molecule.

36. The Fc fusion protein composition according to claim 30, wherein the cell is a mouse myeloma cell.

37. The Fc fusion protein composition according to claim 36, wherein the mouse myeloma cell is NS0 cell or SP2/0-Ag14 cell.

38. The Fc fusion protein composition according to claim 30, wherein the cell is selected from the group consisting of the following (a) to (g):

(a) a CHO cell derived from a Chinese hamster ovary tissue;

(b) a rat myeloma cell line YB2/3HL.P2.G11.16Ag.20 line;

(c) a BHK cell derived from a Syrian hamster kidney tissue;

(d) hybridoma cell;

(e) a human leukemic cell line Namalwa cell;

(f) an embryonic stem cell;

(g) a fertilized egg cell.

42. The Fc fusion protein composition according to claim 30, wherein the Fc fusion protein composition is Fc-fused fibroblast growth factor-8.

43. A cell which produces the Fc fusion protein composition according to claim 30.

45. The cell according to claim 43, which is a mouse myeloma cell.

46. The cell according to claim 45, wherein the mouse myeloma cell is NS0 cell or SP2/0-Ag14 cell.

47. The cell according to claim 43, which is selected from the group consisting of the following (a) to (g):

(a) a CHO cell derived from a Chinese hamster ovary tissue;

(b) a rat myeloma cell line YB2/3HL.P2.G11.16Ag.20 line;

(c) a BHK cell derived from a Syrian hamster kidney tissue;

(d) an antibody-producing hybridoma cell;

(e) a human leukemic cell line Namalwa cell;

(f) an embryonic stem cell;

(g) a fertilized egg cell.

39. An Fc fusion protein composition comprisng an Fc fusion protein having an complex N-glycoside-linked sugar chain at the Fc region of an antibody molecule, wherein the ratio of a sugar chain in which fucose is not bonded to N-acetylglucosamine in the reducing end in the sugar chain is 20% or more of total complex N-glycoside-linked sugar chains which are bound to the Fc region in the composition.

40. The Fc fusion protein composition according to claim 39, wherein the sugar chain in which fucose is not bound is a sugar chain in which 1-position of fucose is not bound to 6-position of N-acetylglucosamine in the reducing end in the complex N-glycoside-linked sugar chain through α

-bond.

41. The Fc fusion protein composition according to claim 39, wherein the antibody molecule belongs to an IgG class.

48. A process for producing an Fc fusion protein