Nucleic acids encoding antibodies to IL-6, and recombinant production of anti-IL-6 antibodies
View Patent ↗The present invention is directed to antibodies and fragments thereof and humanized versions thereof having binding specificity for IL-6. Another embodiment of this invention relates to the antibodies described herein, and binding fragments thereof, comprising the sequences of the V H , V L and CDR polypeptides described herein, and the polynucleotides encoding them. The invention also contemplates conjugates of anti-IL-6 antibodies and binding fragments thereof conjugated to one or more functional or detectable moieties. The invention also contemplates methods of making said anti-IL-6 antibodies and binding fragments thereof. Embodiments of the invention also pertain to the use of anti-IL-6 antibodies, and binding fragments thereof, for the diagnosis, assessment and treatment of diseases and disorders associated with IL-6. These antibodies may bind at least one of soluble IL-6, cell surface expressed IL-6, IL-6/IL-6R and/or prevent the association of IL-6 and IL-6R, the association of IL-6/IL-6R and gp130 and or the formation of IL-6/IL-6R/gp130 multimers and thereby inhibit a biological effect associated with any of the foregoing.
1. At least one isolated or recombinant nucleic acid encoding a V H and/or V L antibody polypeptide which is specific to human IL-6 wherein said V H polypeptide contains CDR1, CDR2 and CDR3 polypeptides which respectively are contained in SEQ ID NO:7, 8 or 659 and 9 and said V L polypeptide contains CDR1, CDR2 and CDR3 polypeptides which respectively are contained in SEQ ID NO: 4, 5 and 6.
2. The at least one isolated or recombinant nucleic acid of claim 1 which encodes a V H antibody polypeptide that is at least 90% identical to the V H antibody polypeptide in SEQ ID NO:656 or 657.
3. A vector containing at least one isolated or recombinant nucleic acid according to claim 2 .
4. A recombinant cell containing at least one isolated or recombinant nucleic acid according to claim 2 or one or more vectors containing said at least one isolated or recombinant nucleic acid.
5. A method of producing a V H and/or V L antibody polypeptide which are specific to human IL-6 said method comprising culturing a cell according to claim 4 under conditions that provide for expression of said V H and/or V L antibody polypeptide.
6. The method of claim 5 wherein the host cell used for expression comprises a polyploid Pichia yeast culture that stably expresses and secretes into the culture medium at least 10-25 mg/liter of said antibody polypeptide or polypeptides.
7. The at least one isolated or recombinant nucleic acid of claim 1 which encodes a V L antibody polypeptide that is at least 90% identical to the V L antibody polypeptide in SEQ ID NO:651 and retains SEQ ID NOs: 7, 8 or 659, and 9.
8. A vector containing at least one isolated or recombinant nucleic acid according to claim 7 .
9. A recombinant cell containing at least one isolated or recombinant nucleic acid according to claim 7 or one or more vectors containing said at least one isolated or recombinant nucleic acid.
10. A method of producing a V H and/or V L antibody polypeptide which are specific to human IL-6 said method comprising culturing a cell according to claim 9 under conditions that provide for expression of said V H and/or V L antibody polypeptide.
11. The at least one isolated or recombinant nucleic acid of claim 1 which encodes a V H antibody polypeptide that is at least 90% identical to the V H antibody polypeptide in SEQ ID NO:656 or 657 and which further encodes a V L antibody polypeptide that is at least 90% identical to the V L antibody polypeptide in SEQ ID NO:651 and retains SEQ ID NOs: 4, 5 and 6.
12. A vector containing at least one isolated or recombinant nucleic acid according to claim 11 .
13. A recombinant cell containing at least one isolated or recombinant nucleic acid according to claim 11 or one or more vectors containing said at least one isolated or recombinant nucleic acid.
14. A method of producing a V H and/or V L antibody polypeptide which are specific to human IL-6 said method comprising culturing a cell according to claim 13 under conditions that provide for expression of said V H and/or V L antibody polypeptide.
15. The at least one isolated nucleic acid of claim 1 wherein said V H antibody polypeptide and said V L antibody polypeptide are encoded by the same isolated or recombinant nucleic acid.
16. A vector containing at least one isolated or recombinant nucleic acid according to claim 15 .
17. A recombinant cell containing at least one isolated or recombinant nucleic acid according to claim 15 or one or more vectors containing said at least one isolated or recombinant nucleic acid.
18. A method of producing a V H and/or V L antibody polypeptide which are specific to human IL-6 said method comprising culturing a cell according to claim 17 under conditions that provide for expression of said V H and/or V L antibody polypeptide.
19. The at least one isolated nucleic acid of claim 1 wherein said V H antibody polypeptide and said V L antibody polypeptide are encoded by two different isolated or recombinant nucleic acids.
20. A vector containing at least one isolated or recombinant nucleic acid according to claim 19 .
21. A recombinant cell containing at least one isolated or recombinant nucleic acid according to claim 19 or one or more vectors containing said at least one isolated or recombinant nucleic acid.
22. A method of producing a V H and/or V L antibody polypeptide which are specific to human IL-6 said method comprising culturing a cell according to claim 21 under conditions that provide for expression of said V H and/or V L antibody polypeptide.
23. The at least one isolated or recombinant nucleic acid of claim 1 which encodes a V H antibody polypeptide identical to SEQ ID NO:656 or 657.
24. A vector containing at least one isolated or recombinant nucleic acid according to claim 23 .
25. A recombinant cell containing at least one isolated or recombinant nucleic acid according to claim 23 or a one or more vectors containing said at least one isolated or recombinant nucleic acid.
26. A method of producing a V H and/or V L antibody polypeptide which are specific to human IL-6 said method comprising culturing a cell according to claim 25 under conditions that provide for expression of said V H and/or V L antibody polypeptide.
27. The at least one isolated or recombinant nucleic acid of claim 1 which encodes a V L antibody polypeptide identical to SEQ ID NO:651.
28. A vector containing at least one isolated or recombinant nucleic acid according to claim 27 .
29. A recombinant cell containing at least one isolated or recombinant nucleic acid according to claim 27 or one or more vectors containing said at least one isolated or recombinant nucleic acid.
30. A method of producing a V H and/or V L antibody polypeptide which are specific to human IL-6 said method comprising culturing a cell according to claim 29 under conditions that provide for expression of said V H and/or V L antibody polypeptide.
31. The at least one isolated or recombinant nucleic acid of claim 1 which encodes a V H antibody polypeptide identical to SEQ ID NO:656 or 657 and which further encodes a V L antibody polypeptide identical to the polypeptide in SEQ ID NO:651.
32. A vector containing at least one isolated or recombinant nucleic acid according to claim 31 .
33. A recombinant cell containing at least one isolated or recombinant nucleic acid according to claim 31 or one or more vectors containing said at least one isolated or recombinant nucleic acid.
34. A method of producing a V H and/or V L antibody polypeptide which are specific to human IL-6 said method comprising culturing a cell according to claim 33 under conditions that provide for expression of said V H and/or V L antibody polypeptide.
35. A vector containing at least one isolated or recombinant nucleic acid according to claim 1 .
36. The vector of claim 35 which is a plasmid or recombinant viral vector.
37. A recombinant cell containing at least one isolated or recombinant nucleic acid according to claim 1 or one or more vectors containing said at least one isolated or recombinant nucleic acid.
38. A method of producing a V H and/or V L antibody polypeptide which is specific to human IL-6 said method comprising culturing a cell according to claim 37 under conditions that provide for expression of said V H and/or V L antibody polypeptide.
39. A recombinant cell according to claim 37 which is selected from a mammalian, yeast, bacterial, and insect cell.
40. The cell of claim 39 which is a yeast cell.
41. The cell of claim 40 which is a diploidal yeast cell.
42. The yeast cell of claim 41 which is a Pichia yeast.
43. The at least one isolated or recombinant nucleic acid of claim 1 which encodes a chimeric V H and/or V L polypeptide.
44. The at least one isolated or recombinant nucleic acid of claim 1 which encodes a humanized V H and/or V L polypeptide.
45. The at least one isolated or recombinant nucleic acid of claim 1 which encodes a chimeric V H and/or V L polypeptide containing a human F c .
46. The at least one isolated or recombinant nucleic acid of claim 45 , wherein said human F c is derived from IgG1, IgG2, IgG3, IgG4, IgG5, IgG6, IgG7, IgG8, IgG9, IgG10, IgG11, IgG12, IgG13, IgG14, IgG15, IgG16, IgG17, IgG18 or IgG19.
47. The at least one isolated or recombinant nucleic acid of claim 1 which encodes for an antibody fragment selected from an F ab fragment, an F ab′ fragment, or an F (ab′)2 fragment.
48. The at least one isolated or recombinant nucleic acid of claim 1 which further encodes for or is attached to an effector moiety.
49. The at least one isolated or recombinant nucleic acid of claim 48 wherein the effector moiety is a detectable moiety or a functional moiety.
50. The at least one isolated or recombinant nucleic acid of claim 49 wherein the detectable moiety is a fluorescent dye, an enzyme, a substrate, a bioluminescent material, a radioactive material, or a chemiluminescent material.
51. The at least one isolated or recombinant nucleic acid of claim 49 wherein the functional moiety is streptavidin, avidin, biotin, a cytotoxin, a cytotoxic agent, or a radioactive material.
52. A nucleic sequence which encodes for the VH region of an anti-IL-6 antibody or antibody fragment according to claim 1 .
53. A nucleic sequence which encodes for the VL region of an anti-IL-6 antibody or antibody fragment according to claim 1 .
54. A vector containing a nucleic acid sequence according to claim 52 .
55. A vector containing a nucleic acid sequence according to claim 53 .