HIGHLY GALACTOSYLATED ANTIBODIES

• US 20140296490A1
• Filed: 08/10/2012
• Published: 10/02/2014
• Est. Priority Date: 08/10/2011
• Status: Abandoned Application

• Current Assignee: Laboratoire francais du Fractionnement et des Biotechnologies
• Original Assignee: Laboratoire francais du Fractionnement et des Biotechnologies
• Inventors: Faid, Valegh, Chevreux, Guillaume
• Application Number:
  US14/237,751
• Publication Number:
  US 20140296490A1
• Time in Patent Office: Days
• US Class Current: 530/387.3
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: C07K 16/04 from milk
• CPC Class Codes: C07K 16/2887 against CD20
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• 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 2317/734 Complement-dependent cytotoxicity [CDC]

Abstract

In one aspect, the disclosure relates to antibodies that are highly galactosylated, methods of production of these antibodies and methods of use of these antibodies.

Claims

1. A composition, comprising:

a population of antibodies wherein the population of antibodies is highly galactosylated.

2-6. -6. (canceled)

7. A composition, comprising:

a population of antibodies,

wherein the level of galactosylation of the antibodies in the population is at least 70%,

wherein the population of antibodies comprises antibodies that comprise mono-galactosylated N-glycans, and

wherein antibodies in the population are encoded by the same nucleic acid sequence.

8. The composition of claim 7, wherein the level of galactosylation of the antibodies in the population is at least 80%, or at least 90%.

9-11. -11. (canceled)

12. A composition, comprising:

a population of antibodies,

wherein the ratio of the level of galactosylation of the antibodies in the population to the level of fucosylation of the antibodies in the population is between 0.8 and 1.2,

wherein the population of antibodies comprises antibodies that comprise mono-galactosylated N-glycans, and

wherein antibodies in the population are encoded by the same nucleic acid sequence.

13. The composition of claim 12, wherein the population of antibodies comprises antibodies that comprise bi-galactosylated N-glycans or antibodies that comprise both mono-galactosylated N-glycans and bi-galactosylated N-glycans.

15. The composition of claim 12, wherein at least 35% of the antibodies in the population comprise bi-galactosylated N-glycans and at least 5% of the antibodies in the population comprise mono-galactosylated N-glycans.

16. The composition of claim 12, wherein the antibodies in the population are transgenically produced in mammary gland epithelial cells.

21. The composition of claim 12, wherein the antibodies of the population of antibodies have an increased level of complement dependent cytotoxicity (CDC) activity when compared to a population of antibodies not produced in mammary gland epithelial cells.

25. The composition of claim 12, wherein the antibodies of the population of antibodies have an increased level of the antibody-dependent cellular cytotoxicity (ADCC) activity when compared to a population of antibodies not produced in mammary gland epithelial cells.

32. The composition of claim 12, wherein the antibodies in the population are anti-CD20 antibodies.

73. Mammary gland epithelial cells that express the population of antibodies of claim 12.

74. A transgenic non-human mammal comprising the mammary gland epithelial cells of claim 73.

14. (canceled)

17-20. -20. (canceled)

22-24. -24. (canceled)

26-31. -31. (canceled)

33-38. -38. (canceled)

39. A method for producing a highly galactosylated population of antibodies, comprising:

producing the population of antibody in mammary gland epithelial cells such that a highly galactosylated population of antibodies is produced.

40. The method of claim 39, wherein the method further comprises collecting the population of antibodies produced.

42. The method of claim 39, wherein the method further comprises determining the CDC activity of the population of antibodies.

47. The method of claim 39, wherein the mammary gland epithelial cells are in a non-human mammal engineered to express a nucleic acid that comprises a sequence that encodes the antibody in its mammary gland.

50. The method of claim 39, wherein the level of galactosylation of the antibodies in the population is at least 70%, at least 80% or at least 90%.

53. The method of claim 39, wherein the level of fucosylation of the antibodies in the population is at least 80%, or at least 90%.

55. The method of claim 39, wherein the ratio of the level of galactosylation of the antibodies in the population to the level of fucosylation of the antibodies in the population is between 0.8 and 1.2.

56. The method of claim 39, wherein the population of antibodies comprises antibodies that comprise mono-galactosylated N-glycans, bi-galactosylated N-glycans, or both mono-galactosylated N-glycans and bi-galactosylated N-glycans.

71. The method of claim 39, wherein the antibodies in the population are anti-CD20 antibodies.

41. (canceled)

43-46. -46. (canceled)

48-49. -49. (canceled)

51-52. -52. (canceled)

54. (canceled)

57-70. -70. (canceled)

72. (canceled)