PROCESS FOR THE PURIFICATION OF RECOMBINANT POLYPEPTIDES
The present invention is directed to a novel process for purifying a recombinant polypeptide from a solution comprising one or more impurities, wherein the process is a chromatography process which uses saccharin. The present invention also provides the use of saccharin in a process for purifying a recombinant polypeptide from a solution comprising one or more impurities, wherein the process is a chromatography process. The invention further provides a wash buffer for purifying using chromatography a recombinant polypeptide from a solution comprising one or more impurities, wherein the wash buffer comprises saccharin.
1 . A process for purifying a recombinant polypeptide from a solution comprising one or more impurities, wherein the process is a chromatography process which comprises the addition of saccharin, wherein the saccharin concentration is from about 0.01 M to about 4 M, and wherein the saccharin is not used to displace the recombinant polypeptide from a chromatography support.
2 . The process according to claim 1 , wherein the chromatography process comprises at least one of: affinity chromatography; ion exchange chromatography; anion exchange chromatography; cation exchange chromatography; gel-permeation or gel filtration chromatography; dye-ligand chromatography; hydrophobic interaction chromatography (HIC); mixed mode chromatography (MMC); and ceramic hydroxyapatite chromatography.
3 . The process according to claim 1 , wherein the process comprises at least one of: (a) a loading step; and, (b) a washing step.
4 . The process according to claim 3 , wherein the process comprises one or more chromatography steps (a), (b):
(a) a loading step comprising a loading solution comprising the recombinant polypeptide and one or more impurities, wherein loading buffer is applied onto a chromatography support;
(b) a washing step comprising a washing buffer wherein the washing buffer is applied onto the chromatography support; and further comprises (c):
(c) an elution step comprising an elution buffer wherein the elution buffer is applied onto the chromatography support.
5 . The process according to claim 1 wherein the chromatography process is Protein A affinity chromatography.
6 . The process according to claim 1 wherein the solution comprising the recombinant polypeptide and one or more impurities, is a cell culture feedstream, or a clarified cell culture fluid.
7 . The process according to claim 1 wherein the recombinant polypeptide is an antigen binding protein.
8 . The process according to claim 7 wherein the antigen binding protein is an antibody.
9 . The process according to claim 1 wherein the one or more impurities are derived from a mammalian cell.
10 . The process according to claim 9 wherein the mammalian cell is selected from the group consisting of: CHO (Chinese Hamster Ovary); NS0; Sp2/0; COS; K562; BHK; PER.C6; and, HEK cells.
11 . The process according to claim 1 wherein the one or more impurities in the process are one or more of: host cell proteins (HCPs), nucleic acids, endotoxins, and cell culture media associated impurities.
12 . The process according to claim 1 wherein the saccharin concentration is from about 0.01 M to about 3.0 M; from about 0.05 M to about 1.0 M; or from about 0.3 M to about 1.5 M.
13 . The process according to claim 4 wherein one or more of the solution, a loading buffer, a washing buffer, and an elution buffer; any one or more of which further comprises one or more of arginine; caprylate; lysine; and, sodium acetate.
14 . The process according to claim 1 wherein saccharin is added to one or more of the solution, loading buffer, and washing buffer.
15 . The process according to claim 14 wherein saccharin is in the form of 2-sulfobenzoic acid ammonium salt; saccharin sodium salt dihydrate; or saccharin sodium salt hydrate.
16 . The process according to claim 1 wherein the purified recombinant polypeptide is further purified.
17 . The process according to claim 1 wherein the purified recombinant polypeptide is formulated to a final therapeutic formulation.
18 . A wash or load buffer for purifying using chromatography a recombinant polypeptide from a solution comprising one or more impurities, wherein the wash or load buffer comprises saccharin, and wherein the saccharin concentration is from about 0.01 M to about 4 M.
19 . A cell culture feedstream comprising a recombinant polypeptide and one or more impurities, wherein the feedstream is a solution comprising saccharin, and wherein the saccharin concentration is from about 0.01 M to about 4 M.
20 . (canceled)
21 . A process for purifying an antibody from a solution comprising the antibody and one or more impurities derived from a mammalian cell, the process comprising a chromatography process comprising a chromatography support and:
(a) loading the solution comprising the antibody and one or more impurities derived from a mammalian cell on to the chromatography support by applying the solution and a loading buffer to the chromatography support;
(b) washing the chromatography support by applying a washing buffer to the chromatography support;
(c) eluting the antibody from the chromatography support by applying an elution buffer to the chromatography support;
wherein one or more of the loading buffer and the washing buffer comprise from about 0.3 M to about 1.5 M saccharin.
22 . The process of claim 21 wherein:
(1) the chromatography process is Protein A chromatography; and,
(2) one or more of the solution, the loading buffer, the washing buffer, and the elution buffer; further comprises one or more of arginine, caprylate, lysine, and, sodium acetate.