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 liquid pharmaceutical composition comprising adalimumab and one or more of sorbitol, a sugar, glycerol, or a preservative,
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 no greater than 1.3 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 liquid composition of claim 1 , wherein the composition is packaged in a pre-filled syringe.
3. The liquid composition of claim 2 , wherein the composition comprises 50 mg/ml of adalimumab.
4. The liquid composition of claim 3 , wherein the composition is suitable for subcutaneous injection.
5. The liquid composition of claim 4 , wherein the composition comprises sorbitol.
6. The liquid composition of claim 4 , wherein the composition comprises glycerol.
7. The liquid composition of claim 4 , wherein the composition comprises a sugar.
8. The liquid composition of claim 4 , wherein the composition comprises a preservative.
9. The liquid composition of claim 1 , wherein the cathepsin L activity is no greater than 0.85 RFU/s/mg of adalimumab.
10. The liquid composition of claim 9 , wherein the composition is packaged in a pre-filled syringe, wherein the composition comprises 50 mg/ml of adalimumab, and wherein the composition is suitable for subcutaneous injection.
11. The liquid composition of claim 10 , wherein the composition comprises sorbitol.
12. The liquid composition of claim 10 , wherein the composition comprises glycerol.
13. The liquid composition of claim 10 , wherein the composition comprises a sugar.
14. The liquid composition of claim 10 , wherein the composition comprises a preservative.
15. The liquid composition of claim 1 , wherein the cathepsin L activity is no greater than 1.0 RFU/s/mg of adalimumab.
16. The liquid composition of claim 15 , wherein the composition is packaged in a pre-filled syringe, wherein the composition comprises 50 mg/ml of adalimumab, and wherein the composition is suitable for subcutaneous injection.
17. The liquid composition of claim 16 , wherein the composition comprises sorbitol.
18. The liquid composition of claim 16 , wherein the composition comprises glycerol.
19. The liquid composition of claim 16 , wherein the composition comprises a sugar.
20. The liquid composition of claim 16 , wherein the composition comprises a preservative.
21. The liquid composition of claim 1 , wherein the cathepsin L activity is no greater than 0.6 RFU/s/mg of adalimumab.
22. The liquid composition of claim 21 , wherein the composition is packaged in a pre-filled syringe, wherein the composition comprises 50 mg/ml of adalimumab, and wherein the composition is suitable for subcutaneous injection.
23. The liquid composition of claim 22 , wherein the composition comprises sorbitol.
24. The liquid composition of claim 22 , wherein the composition comprises glycerol.
25. The liquid composition of claim 22 , wherein the composition comprises a sugar.
26. The liquid composition of claim 22 , wherein the composition comprises a preservative.
27. The liquid composition of claim 4 , wherein the diluted composition has an adalimumab concentration of 20 μg/ml.
28. The liquid composition of claim 4 , wherein the diluted composition has an adalimumab concentration of 50 μg/ml.
29. The liquid composition of claim 4 , wherein step i) of the cathepsin L kinetic assay comprises diluting the composition 600 fold.
30. The liquid composition of claim 1 , wherein the composition comprises 50 mg/ml of adalimumab, and wherein the composition is suitable for subcutaneous injection.