IL-18 binding protein (IL-18BP) and antibodies in inflammatory diseases
The present invention provides means and methods for treating Interleukin 18 (IL-18)-associated diseases and disorders. In particular, the present invention discloses antibodies specific for free IL-18 and IL-18 Binding Protein (IL-18BP) for use in such treatments and for the diagnosis of the diseases and disorders.
1. A composition comprising a pharmaceutically acceptable carrier and a recombinant IL-18 binding protein (IL-18BP) having the amino acid sequence of SEQ ID NO: 7, and at least one deletion variant comprising a deletion of 1 to 5 amino acid residues at the C-terminal end of the IL-18BP, wherein the proportion of the at least one deletion variant in the composition is less than 30%.
2. The composition of claim 1 , wherein the proportion of the at least one deletion variant in the composition is less than 20%.
3. The composition of claim 2 , wherein the proportion of the at least one deletion variant in the composition is less than 15%.
4. The composition of claim 3 , wherein the proportion of the at least one deletion variant in the composition is less than 10%.
5. The composition of claim 4 , wherein the proportion of the at least one deletion variant in the composition is less than 7.5%.
6. The composition of claim 5 , wherein the proportion of the at least one deletion variant in the composition is less than 5%.
7. The composition of claim 6 , wherein the proportion of the at least one deletion variant in the composition is less than 2.5%.
8. The composition of claim 7 , wherein the proportion of the at least one deletion variant in the composition is less than 1%.
9. The composition of claim 1 , wherein the composition comprises sodium chloride, sodium dihydrogen phosphate monohydrate, disodium phosphate dehydrate, and water.
10. The composition of claim 1 , wherein the recombinant IL-18BP binds to IL-18 with a binding affinity of 20-50 pM based on BIAcore measurements.
11. A method of preparing the composition of claim 1 , comprising growing a recombinant Chinese Hamster Ovary (CHO) cell under conditions to express the recombinant IL-18BP, and purifying the recombinant IL-18BP and the at least one deletion variant from the CHO cell.
12. The method of claim 11 , wherein the recombinant IL-18BP and the at least one deletion variant are purified by a method comprising: (a) removing the recombinant CHO cells and cell debris from a cell culture supernatant by centrifugation, and/or diafiltration; (b) concentrating the harvest containing the recombinant IL-18BP and the at least one deletion variant; (c) capturing the recombinant IL-18BP and the at least one deletion variant on an anion-exchange resin to remove salts and cell culture nutrients; (d) eluting the recombinant IL-18BP and the at least one deletion variant; and optionally (e) applying additional chromatographic steps to further purifying the recombinant IL-18BP and the at least one deletion variant.
13. A composition comprising a pharmaceutically acceptable carrier and a recombinant IL-18 binding protein (IL-18BP) having the amino acid sequence of SEQ ID NO: 7, and at least one deletion variant comprising a deletion of 1 to 30 amino acid residues at the N-terminal end of the IL-18BP, wherein the proportion of the at least one deletion variant in the composition is less than 30%.
14. The composition of claim 13 , wherein the proportion of the at least one deletion variant in the composition is less than 20%.
15. The composition of claim 14 , wherein the proportion of the at least one deletion variant in the composition is less than 15%.
16. The composition of claim 15 , wherein the proportion of the at least one deletion variant in the composition is less than 10%.
17. The composition of claim 16 , wherein the proportion of the at least one deletion variant in the composition is less than 7.5%.
18. The composition of claim 17 , wherein the proportion of the at least one deletion variant in the composition is less than 5%.
19. The composition of claim 18 , wherein the proportion of the at least one deletion variant in the composition is less than 2.5%.
20. The composition of claim 19 , wherein the proportion of the at least one deletion variant in the composition is less than 1%.
21. The composition of claim 13 , wherein the composition comprises sodium chloride, sodium dihydrogen phosphate monohydrate, disodium phosphate dehydrate, and water.
22. The composition of claim 13 , wherein the recombinant IL-18BP binds to IL-18 with a binding affinity of 20-50 pM based on BIAcore measurements.
23. A method of preparing the composition of claim 13 , comprising growing a recombinant Chinese Hamster Ovary (CHO) cell under conditions to express the recombinant IL-18BP, and purifying the recombinant IL-18BP and the at least one deletion variant from the CHO cell.
24. The method of claim 23 , wherein the recombinant IL-18BP and the at least one deletion variant are purified by a method comprising: (a) removing the recombinant CHO cells and cell debris from a cell culture supernatant by centrifugation, and/or diafiltration; (b) concentrating the harvest containing the recombinant IL-18BP and the at least one deletion variant; (c) capturing the recombinant IL-18BP and the at least one deletion variant on an anion-exchange resin to remove salts and cell culture nutrients; (d) eluting the recombinant IL-18BP and the at least one deletion variant; and optionally (e) applying additional chromatographic steps to further purifying the recombinant IL-18BP and the at least one deletion variant.
25. A composition comprising a pharmaceutically acceptable carrier and a recombinant IL-18 binding protein (IL-18BP) having the amino acid sequence of SEQ ID NO: 7, at least one first deletion variant comprising a deletion of 1 to 5 amino acid residues at the C-terminal end of the IL-18BP, and at least one second deletion variant comprising a deletion of 1 to 30 amino acid residues at the N-terminal end of the IL-18BP, wherein the proportion of the at least one first deletion variant and the at least one second deletion variant in the composition is less than 30%.
26. The composition of claim 25 , wherein the proportion of the at least one first deletion variant and the at least one second deletion variant in the composition is less than 20%.
27. The composition of claim 26 , wherein the proportion of the at least one first deletion variant and the at least one second deletion variant in the composition is less than 15%.
28. The composition of claim 27 , wherein the proportion of the at least one first deletion variant and the at least one second deletion variant in the composition is less than 10%.
29. The composition of claim 28 , wherein the proportion of the at least one first deletion variant and the at least one second deletion variant in the composition is less than 7.5%.
30. The composition of claim 29 , wherein the proportion of the at least one first deletion variant and the at least one second deletion variant in the composition is less than 5%.
31. The composition of claim 25 , wherein the composition comprises sodium chloride, sodium dihydrogen phosphate monohydrate, disodium phosphate dehydrate, and water.
32. The composition of claim 25 , wherein the recombinant IL-18BP binds to IL-18 with a binding affinity of 20-50 pM based on BIAcore measurements.
33. A method of preparing the composition of claim 25 , comprising growing a recombinant Chinese Hamster Ovary (CHO) cell under conditions to express the recombinant IL-18BP, and purifying the recombinant IL-18BP, the at least one first deletion variant and the at least one second deletion variant from the CHO cell.
34. The method of claim 33 , wherein the recombinant IL-18BP, the at least one first deletion variant, and the at least one second deletion variant are purified by a method comprising: (a) removing the recombinant CHO cells and cell debris from a cell culture supernatant by centrifugation, and/or diafiltration; (b) concentrating the harvest containing the recombinant IL-18BP, the at least one first deletion variant, and the at least one second deletion variant; (c) capturing the recombinant IL-18BP, the at least one first deletion variant, and the at least one second deletion variant on an anion-exchange resin to remove salts and cell culture nutrients; (d) eluting the recombinant IL-18BP, the at least one first deletion variant, and the at least one second deletion variant; and optionally (e) applying additional chromatographic steps to further purifying the recombinant IL-18BP, the at least one first deletion variant and the at least one second deletion variant.
35. The method of claim 34 , wherein the recombinant IL-18BP, the at least one first deletion variant, and the at least one second deletion variant are purified by a method comprising: (a) removing the recombinant CHO cells and cell debris from a cell culture supernatant by centrifugation, and/or diafiltration; (b) concentrating the harvest containing the recombinant IL-18BP, the at least one first deletion variant, and the at least one second deletion variant and diafiltrating against a basic borate buffer at a pH higher than pH 7.5; (c) capturing the recombinant IL-18BP, the at least one first deletion variant, and the at least one second deletion variant on a TMAE Hi-Cap anion-exchange resin to remove salts and cell culture nutrients; (d) eluting the recombinant IL-18BP, the at least one first deletion variant, and the at least one second deletion variant with a borate buffer containing NaCl; and optionally (e) applying additional chromatographic steps to further purifying the recombinant IL-18BP, the at least one first deletion variant, and the at least one second deletion variant.