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 of treating a disorder in which TNFα activity is detrimental in a subject, the method comprising administering a composition comprising a therapeutically effective amount of adalimumab to the subject such that the disorder is treated,
wherein the adalimumab is produced in a Chinese Hamster Ovary (CHO) cell expression system;
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; and
wherein the composition is characterized in that when the composition is assayed in a cathepsin L kinetic assay, a level of cathepsin L activity from 0.6 to 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 0.6 to 1.0 RFU/s/mg of adalimumab.
3. The method of claim 1 , wherein the cathepsin L activity is from 0.6 to 1.3 RFU/s/mg of adalimumab.
4. A method of treating a disorder in which TNFα activity is detrimental in a subject, the method comprising administering a composition comprising a therapeutically effective amount of adalimumab to the subject such that the disorder is treated,
wherein the adalimumab is produced in a Chinese Hamster Ovary (CHO) cell expression system;
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; and
wherein the composition is characterized in that when the composition is assayed in a cathepsin L kinetic assay, a level of cathepsin L activity from 0.4 to 0.9 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.
5. The method of claim 4 , wherein the cathepsin L activity is from 0.4 to 0.6 RFU/s/mg of adalimumab.
6. The method of claim 4 , wherein the cathepsin L activity is from 0.4 to 0.85 RFU/s/mg of adalimumab.
7. The method of any one of claims 1 and 2 - 6 , wherein the composition is packaged in a pre-filled syringe.
8. The method of any one of claims 1 and 2 - 6 , wherein the composition comprises 50 mg/ml of adalimumab.
9. The method of any one of claims 1 and 2 - 6 , wherein the composition is suitable for subcutaneous injection.
10. The method of any one of claims 1 and 2 - 6 , wherein the diluted composition has an adalimumab concentration of 20 μg/ml.
11. The method of any one of claims 1 and 2 - 6 , wherein the diluted composition has an adalimumab concentration of 50 μg/ml.
12. The method of any one of claims 1 and 2 - 6 , wherein step i) of the cathepsin L kinetic assay comprises diluting the composition 600 fold.
13. The method of claim 4 , wherein the disorder is rheumatoid arthritis.
14. The method of claim 4 , wherein the disorder is Crohn's disease.
15. The method of claim 4 , wherein the disorder is ulcerative colitis.
16. The method of claim 4 , wherein the disorder is ankylosing spondylitis.
17. The method of claim 4 , wherein the disorder is psoriatic arthritis.
18. The method of claim 4 , wherein the disorder is psoriasis.
19. The method of claim 4 , wherein the disorder is juvenile rheumatoid arthritis.