Purified antibody composition
The invention provides a method for producing a host cell protein-(HCP) reduced antibody preparation from a mixture comprising an antibody and at least one HCP, comprising an ion exchange separation step wherein the mixture is subjected to a first ion exchange material, such that the HCP-reduced antibody preparation is obtained.
1. A method for preparing an adalimumab formulation, comprising the step of mixing a liquid composition comprising adalimumab with a pharmaceutically acceptable carrier, wherein the adalimumab is expressed in a Chinese Hamster Ovary (CHO) cell expression system, and the composition is characterized in that when the composition is assayed in a cathepsin L kinetic assay, a level of cathepsin L activity of less than 1.84 RFU/s/mg of adalimumab is observed, wherein the cathepsin L kinetic assay comprises:
i) diluting the composition in a polystyrene container in a solution containing 25 mM NaOAc, 5 mM DTT and 1 mM EDTA at pH 5.5,
ii) adding dextran sulfate to a concentration of 0.035 μg/mL and incubating at 37° C. for six hours,
iii) adding Z-leucine-arginine covalently bound at its C-terminus to a fluorescent 7-amino-4-methyl coumarin (Z-leucine-arginine-AMC), wherein the diluting, adding, and incubating steps are sufficient to permit the measurement of cathepsin L hydrolysis of the Z-leucine-arginine-AMC within a linear range, and
iv) measuring Z-leucine-arginine-AMC hydrolysis in the linear range in RFU/s/mg of adalimumab.
2. The method of claim 1 , wherein the cathepsin L activity is from 0.4 to no greater than 1.0 RFU/s/mg of adalimumab.
3. The method of claim 1 , wherein the cathepsin L activity is from 0.4 to no greater than 1.3 RFU/s/mg of adalimumab.
4. The method of claim 1 , wherein the cathepsin L activity is from 0.4 to no greater than 0.6 RFU/s/mg of adalimumab.
5. The method of claim 1 , wherein the cathepsin L activity is from 0.4 to no greater than 0.85 RFU/s/mg of adalimumab.
6. The method of claim 1 , wherein the cathepsin L activity is from 0.4 to no greater than 0.9 RFU/s/mg of adalimumab.
7. The method of claim 1 , wherein the cathepsin L activity is from 0.5 to 1.5 RFU/s/mg of adalimumab.
8. The method of claim 1 , wherein the cathepsin L activity is from 0.4 to less than 1.84 RFU/s/mg of adalimumab.
9. The method of claim 1 , wherein the formulation is packaged in a pre-filled syringe.
10. The method of claim 1 , wherein the formulation comprises 50 mg/ml of adalimumab.
11. The method of claim 1 , wherein the formulation is suitable for subcutaneous injection.
12. The method of claim 1 , wherein the diluted composition has an adalimumab concentration of 20 μg/ml.
13. The method of claim 1 , wherein the diluted composition has an adalimumab concentration of 50 μg/ml.
14. The method of claim 1 , wherein step i) of the cathepsin L kinetic assay comprises diluting the composition 600 fold.
15. The method of claim 1 , wherein the pharmaceutically acceptable carrier comprises mannitol.
16. The method of claim 1 , wherein the pharmaceutically acceptable carrier comprises sodium chloride.
17. The method of claim 8 , wherein the formulation is packaged in a pre-filled syringe.
18. The method of claim 8 , wherein the formulation comprises 50 mg/ml of adalimumab.
19. The method of claim 8 , wherein the formulation is suitable for subcutaneous injection.
20. The method of claim 8 , wherein the diluted composition has an adalimumab concentration of 20 μg/ml.
21. The method of claim 8 , wherein the diluted composition has an adalimumab concentration of 50 μg/ml.
22. The method of claim 8 , wherein step i) of the cathepsin L kinetic assay comprises diluting the composition 600 fold.
23. The method of claim 8 , wherein the pharmaceutically acceptable carrier comprises mannitol.
24. The method of claim 8 , wherein the pharmaceutically acceptable carrier comprises sodium chloride.
25. An adalimumab formulation prepared according to the method of claim 1 .
26. An adalimumab formulation prepared according to the method of claim 8 .
27. A method of treating a disorder in which TNFα activity is detrimental in a subject, the method comprising administering to the subject a therapeutically effective amount of the adalimumab formulation of claim 25 such that the disorder is treated, wherein the disorder is selected from the group consisting of rheumatoid arthritis, Crohn's disease, ulcerative colitis, ankylosing spondylitis, psoriatic arthritis, psoriasis, and juvenile rheumatoid arthritis.
28. A method of treating a disorder in which TNFα activity is detrimental in a subject, the method comprising administering to the subject a therapeutically effective amount of the adalimumab formulation of claim 26 such that the disorder is treated, wherein the disorder is selected from the group consisting of rheumatoid arthritis, Crohn's disease, ulcerative colitis, ankylosing spondylitis, psoriatic arthritis, psoriasis, and juvenile rheumatoid arthritis.