Methods for purifying antibodies
The present invention relates to a method of purifying a recombinant polypeptide from Host Cell Proteins (HCP), the method comprising: (a) applying a solution comprising the recombinant polypeptide and HCP to a superantigen chromatography solid support, (b) washing the superantigen chromatography solid support with a wash buffer comprising caprylate and arginine; and (c) eluting the recombinant polypeptide from the superantigen chromatography solid support.
1. A method of purifying a recombinant polypeptide from Host Cell Proteins (HCP), the method comprising: (a) applying a solution comprising the recombinant polypeptide and HCP to a superantigen chromatography solid support, (b) washing the superantigen chromatography solid support with a wash buffer comprising at least about 75 mM caprylate and greater than about 0.5 M arginine; and (c) eluting the recombinant polypeptide from the superantigen chromatography solid support.
2. The method according to claim 1 , wherein the caprylate is sodium caprylate.
3. The method according to claim 1 , wherein the wash buffer comprises about 75 mM to about 300 mM caprylate.
4. The method according to claim 1 , wherein the wash buffer comprises about 0.75 M to about 1.5 M arginine.
5. The method according to claim 1 , wherein the eluted recombinant polypeptide contains less than about 2% fragmented recombinant polypeptide.
6. The method according to claim 1 , wherein the HCP is derived from a mammalian cell.
7. The method according to claim 1 , wherein the HCP is phospholipase B-Like 2 protein.
8. The method according to claim 1 , wherein the HCP is cathepsin L.
9. The method according to claim 8 , wherein the purification of the recombinant polypeptide from cathepsin L is measured by a reduced cathepsin L activity in the eluate of step (c).
10. The method according to claim 1 , wherein the pH of the wash buffer is between pH 7 to pH 9.
11. The method according to claim 1 , wherein the recombinant polypeptide is a monoclonal antibody (mAb).
12. The method according to claim 11 , wherein the mAb is an IgG1, or an IgG4.
13. The method according to claim 1 , wherein the wash buffer does not contain sodium chloride.
14. The method according to claim 1 , wherein the superantigen is selected from the group consisting of Protein A, Protein G, and Protein L.
15. The method according to claim 1 , wherein after step (c) the amount of HCP is less than about 200 ng HCP/mg product.
16. A method of purifying a recombinant polypeptide from phospholipase B-Like 2 protein, the method comprising: (a) applying a solution comprising the recombinant polypeptide and phospholipase B-Like 2 protein to a superantigen chromatography solid support, (b) washing the superantigen chromatography solid support with a wash buffer comprising about 100 mM caprylate and about 1.1 M arginine; and (c) eluting the recombinant polypeptide from the superantigen chromatography solid support.
17. A method of purifying a recombinant polypeptide from cathepsin L, the method comprising: (a) applying a solution comprising the recombinant polypeptide and cathepsin L to a superantigen chromatography solid support, (b) washing the superantigen chromatography solid support with a wash buffer comprising about 150 mM caprylate and about 1.1 M arginine; and (c) eluting the recombinant polypeptide from the superantigen chromatography solid support.