ANTI-ERYTHROPOIETIN ANTIBODIES

• US 20130295113A1
• Filed: 05/07/2013
• Published: 11/07/2013
• Est. Priority Date: 05/07/2012
• Status: Active Grant

• Current Assignee: Amgen Inc.
• Original Assignee: Amgen Inc.
• Inventors: Mytych, Daniel T., Chirmule, Narendra, Weeraratne, Dohan K.
• Granted Patent: US 9,441,039 B2
• Time in Patent Office: Days
• US Class Current: 424/158.1
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: C07K 16/22 against growth factors ; against growth regulators
• CPC Class Codes: C07K 16/26 against hormones ; against hormone releasing or inhibiting factors
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/34 Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: G01N 2800/22 Haematology
• CPC Class Codes: G01N 2800/50 Determining the risk of developing a disease
• CPC Class Codes: G01N 33/6854 Immunoglobulins

Abstract

The present disclosure provides compositions and methods relating to antibodies that specifically bind to human erythropoietin. The disclosure provides nucleic acids encoding such antibodies and methods of making and using such antibodies.

Claims

1. An isolated antibody or fragment of an antibody, wherein the antibody or the fragment specifically binds to human erythropoietin and comprises:

a. a VH CDR1 having an amino acid sequence identical to or comprising 0, 1, 2, or 3 amino acid residue substitutions relative to SEQ ID NO;

36;

b. a VH CDR2 having an amino acid sequence identical to or comprising 0, 1, 2, or 3 amino acid residue substitutions relative to SEQ ID NO;

37;

c. a VH CDR3 having an amino acid sequence identical to or comprising 0, 1, 2, or 3 amino acid residue substitutions relative to SEQ ID NO;

38;

d. a VL CDR1 having an amino acid sequence identical to or comprising 0, 1, 2, or 3 amino acid residue substitutions relative to SEQ ID NO;

6;

e. a VL CDR2 having an amino acid sequence identical to or comprising 0, 1, 2, or 3 amino acid residue substitutions relative to SEQ ID NO;

7; and

f. a VL CDR3 having an amino acid sequence identical to or comprising 0, 1, 2, or 3 amino acid residue substitutions relative to SEQ ID NO;

8;

or

g. a VH CDR1 having an amino acid sequence identical to or comprising 1, 2, or 3 amino acid residue substitutions relative to SEQ ID NO;

42;

h. a VH CDR2 having an amino acid sequence identical to or comprising 1, 2, or 3 amino acid residue substitutions relative to SEQ ID NO;

43;

i. a VH CDR3 having an amino acid sequence identical to or comprising 1, 2, or 3 amino acid residue substitutions relative to SEQ ID NO;

44;

j. a VL CDR1 having an amino acid sequence identical to or comprising 1, 2, or 3 amino acid residue substitutions relative to SEQ ID NO;

12;

k. a VL CDR2 having an amino acid sequence identical to or comprising 1, 2, or 3 amino acid residue substitutions relative to SEQ ID NO;

13; and

l. a VL CDR3 having an amino acid sequence identical to or comprising 1, 2, or 3 amino acid residue substitutions relative to SEQ ID NO;

14;

or

m. a VH CDR1 having an amino acid sequence identical to or comprising 1, 2, or 3 amino acid residue substitutions relative to SEQ ID NO;

48;

n. a VH CDR2 having an amino acid sequence identical to or comprising 1, 2, or 3 amino acid residue substitutions relative to SEQ ID NO;

49;

o. a VH CDR3 having an amino acid sequence identical to or comprising 1, 2, or 3 amino acid residue substitutions relative to SEQ ID NO;

50;

p. a VL CDR1 having an amino acid sequence identical to or comprising 1, 2, or 3 amino acid residue substitutions relative to SEQ ID NO;

18;

q. a VL CDR2 having an amino acid sequence identical to or comprising 1, 2, or 3 amino acid residue substitutions relative to SEQ ID NO;

19; and

r. a VL CDR3 having an amino acid sequence identical to or comprising 1, 2, or 3 amino acid residue substitutions relative to SEQ ID NO;

20;

or

s. a VH CDR1 having an amino acid sequence identical to or comprising 1, 2, or 3 amino acid residue substitutions relative to SEQ ID NO;

54;

t. a VH CDR2 having an amino acid sequence identical to or comprising 1, 2, or 3 amino acid residue substitutions relative to SEQ ID NO;

55;

u. a VH CDR3 having an amino acid sequence identical to or comprising 1, 2, or 3 amino acid residue substitutions relative to SEQ ID NO;

56;

v. a VL CDR1 having an amino acid sequence identical to or comprising 1, 2, or 3 amino acid residue substitutions relative to SEQ ID NO;

24;

w. a VL CDR2 having an amino acid sequence identical to or comprising 1, 2, or 3 amino acid residue substitutions relative to SEQ ID NO;

25; and

x. a VL CDR3 having an amino acid sequence identical to or comprising 1, 2, or 3 amino acid residue substitutions relative to SEQ ID NO;

26;

or

y. a VH CDR1 having an amino acid sequence identical to or comprising 1, 2, or 3 amino acid residue substitutions relative to SEQ ID NO;

60;

z. a VH CDR2 having an amino acid sequence identical to or comprising 1, 2, or 3 amino acid residue substitutions relative to SEQ ID NO;

61;

aa. a VH CDR3 having an amino acid sequence identical to or comprising 1, 2, or 3 amino acid residue substitutions relative to SEQ ID NO;

62;

bb. a VL CDR1 having an amino acid sequence identical to or comprising 1, 2, or 3 amino acid residue substitutions relative to SEQ ID NO;

30;

cc. a VL CDR2 having an amino acid sequence identical to or comprising 1, 2, or 3 amino acid residue substitutions relative to SEQ ID NO;

31; and

dd. a VL CDR3 having an amino acid sequence identical to or comprising 1, 2, or 3 amino acid residue substitutions relative to SEQ ID NO;

32.

3. An isolated antibody which competes for binding with the antibodies of claims 1 or 2.

4. The antibody of claim 3, wherein the antibody is a monoclonal antibody.

5. The antibody of claim 3, wherein the antibody is a chimeric antibody, a humanized antibody, or a fully human antibody.

6. A sterile composition comprising the isolated antibody or antibody fragment of any one of claim 1 or 2.

7. The composition of claim 6 further comprising a pharmaceutically acceptable carrier.

8. An isolated nucleic acid encoding the antibody of claim 1 or 2.

9. A vector comprising the nucleic acid of claim 8.

10. A host cell comprising the vector or nucleic acid of claim 9.

11. A method for producing an antibody comprising culturing the host cell of claim 10 under suitable conditions and recovering said antibody.

2. An isolated antibody or antibody fragment, wherein the antibody or the fragment specifically binds human erythropoietin and comprises:

a heavy chain variable domain having at least 90% identity to the amino acid of SEQ ID NO;

69 and comprises a light chain variable domain having at least 90% identity to the amino acid sequence of SEQ ID NO;

63;

or comprises a heavy chain variable domain having at least 90% identity to the amino acid of SEQ ID NO;

70 and comprises a light chain variable domain having at least 90% identity to the amino acid sequence of SEQ ID NO;

64;

or comprises a heavy chain variable domain having at least 90% identity to the amino acid of SEQ ID NO;

71 and comprises a light chain variable domain having at least 90% identity to the amino acid sequence of SEQ ID NO;

65;

or comprises a heavy chain variable domain having at least 90% identity to the amino acid of SEQ ID NO;

72 and comprises a light chain variable domain having at least 90% identity to the amino acid sequence of SEQ ID NO;

65;

or comprises a heavy chain variable domain having at least 90% identity to the amino acid of SEQ ID NO;

73 and comprises a light chain variable domain having at least 90% identity to the amino acid sequence of SEQ ID NO;

66;

or comprises a heavy chain variable domain having at least 90% identity to the amino acid of SEQ ID NO;

74 and comprises a light chain variable domain having at least 90% identity to the amino acid sequence of SEQ ID NO;

66;

or comprises a heavy chain variable domain having at least 90% identity to the amino acid of SEQ ID NO;

75 and comprises a light chain variable domain having at least 90% identity to the amino acid sequence of SEQ ID NO;

67;

or comprises a heavy chain variable domain having at least 90% identity to the amino acid of SEQ ID NO;

76 and comprises a light chain variable domain having at least 90% identity to the amino acid sequence of SEQ ID NO;

68.

12. A method of measuring anti-human erythropoietin IgG4 antibody levels in a human subject comprising determining in vitro a level of anti-human erythropoietin IgG4 antibodies in a sample from said subject and comparing said level to a level obtained from the anti-human erythropoietin IgG4 antibody 8C10 or 3A4.

18. The method of claims 12-17, wherein said antibody 8C10 or 3A4 is used as a positive control.

19. The method of claims 12-17, wherein said antibody 8C10 or 3A4 is used to generate a standard curve.

13. A method of measuring anti-human erythropoietin IgG4 antibody levels in a human subject comprising:

a. contacting in vitro a blood or serum sample from said subject with a human erythropoietin, wherein the human erythropoietin captures anti-human IgG4 antibodies;

b. contacting in vitro antibody 8C10 or 3A4 with a human erythropoietin, wherein the human erythropoietin captures 8C10 or 3A4;

c. contacting said captured antibodies of steps a) and b) with a secondary anti-human IgG4 antibody that has been detectably labeled;

d. measuring the levels of said secondary antibody in each sample from step c).

14. The method of claim 13, further comprising comparing the levels of the subject sample to the levels of 8C10 or 3A4, wherein an increase in a subject'"'"'s anti-erythropoietin IgG4 levels indicates the onset or risk of amPRCA.

15. A method of detecting antibody-mediated pure red cell aplasia (amPRCA) in a human subject comprising determining in vitro a level of anti-human erythropoietin IgG4 antibodies in a sample from said subject and comparing said level to a level obtained from the anti-human erythropoietin IgG4 antibody 8C10 or 3A4, wherein an increase in a subject'"'"'s anti-erythropoietin IgG4 levels indicates the onset or risk of amPRCA.

16. A method of predicting the onset of antibody-mediated pure red cell aplasia (amPRCA) comprising determining in vitro a level of anti-human erythropoietin IgG4 antibodies in a sample from said subject and comparing said level to a level obtained from the anti-human erythropoietin IgG4 antibody 8C10 or 3A4, wherein an increase in a subject'"'"'s anti-erythropoietin IgG4 levels indicates the onset or risk of amPRCA.

17. A method of predicting the risk of antibody-mediated pure red cell aplasia (amPRCA) comprising determining in vitro a level of anti-human erythropoietin IgG4 antibodies in a sample from said subject and comparing said level to a level obtained from the anti-human erythropoietin IgG4 antibody 8C10 or 3A4, wherein an increase in a subject'"'"'s anti-erythropoietin IgG4 levels indicates the onset or risk of amPRCA.

20. A kit for detecting amPRCA or predicting the risk or onset of amPRCA comprising an anti-human erythropoietin IgG4 antibody and human erythropoietin.

21. The kit of claim 20, wherein said antibody is 8C10 or 3A4.