Claims for Patent: 10,364,268

Summary for Patent: 10,364,268

Title:	Ion exchange membrane chromatography
Abstract:	Methods of enhancing efficiency of downstream chromatography steps for purification of proteins comprising: (a) passing a composition comprising a polypeptide of interest and various contaminants through an ion exchange membrane, wherein the polypeptide and the membrane have opposite charge, at operating conditions comprised of a buffer having a pH sufficiently distinct from the pi of the polypeptide to enhance the charge of the polypeptide and a low ionic strength effective to prevent the shielding of charges by buffer ions, which cause the membrane to bind the polypeptide and at least one contaminant, (b) overloading the ion exchange membrane such that at least one contaminant remains bound to the membrane while the polypeptide of interest is primarily in the effluent; (c) collecting the effluent from the ion exchange membrane comprising the polypeptide of interest; (d) subjecting the membrane effluent comprising the polypeptide of interest to a purification step of similar charge as the previous membrane, and (e) recovering the purified polypeptide from the effluent of the charged ion exchange chromatography purification step.
Inventor(s):	Bill, Jr.; Jerome Joseph (South San Francisco, CA), Brown; Arick Michael (South San Francisco, CA), Dowd; Christopher John (South San Francisco, CA), Thayer; Brooke Ellen (South San Francisco, CA)
Assignee:	Genentech, Inc. (South San Francisco, CA)
Application Number:	14/365,449
Patent Claims:	1. A method of enhancing efficiency of downstream chromatography steps for purification of antibodies consisting of: a. passing a harvested cell culture fluid composition comprising an antibody of interest and various contaminants through a cation exchange membrane, wherein the antibody and the membrane have opposite charge, at operating conditions comprised of a buffer having a pH of about 1 to about 5 pH units below the pl of the antibody and a conductivity of .ltoreq.about 40 mS/cm, which cause the membrane to bind the antibody and at least one ionic polymer contaminant, where in the ionic polymer contaminant is one or more of polyethyleneimine (PEI), polyvinylamine, polyarginine, polyvinylsulfonic acid, or polyacrylic acid; b. overloading the cation exchange membrane to promote competitive adsorption such that at least one ionic polymer contaminant remains bound to the membrane while the antibody of interest is primarily in the effluent; c. collecting the effluent from the cation exchange membrane comprising the antibody of interest; d. subjecting the membrane effluent comprising the antibody of interest to a cation exchange chromatography purification step, wherein the dynamic binding capacity of the cation exchange chromatography purification step is significantly improved as a result of steps (a) through (c); e. recovering the purified antibody from the effluent of the cation exchange chromatography purification step; f. subjecting the cation exchange chromatography purified pool to a final polishing step; and g. recovering the purified antibody of interest. 2. The method of claim 1, wherein the harvested cell culture fluid composition comprising the antibody of interest and various contaminants comprises E. Coli Proteins (ECP). 3. The method of claim 1 wherein the pH is about 1 to about 4 pH units below the pl of the antibody. 4. The method of claim 1 wherein the pH is about 1 to about 3 pH units below the pl of the antibody. 5. The method of claim 1 wherein the pH is about 1 to about 2 pH units below the pI of the antibody. 6. The method of claim 1 wherein the pH is about 1 pH unit below the pl of the antibody. 7. The method of claim 1 wherein the conductivity is .ltoreq.about 20 mS/cm. 8. The method of claim 1 wherein the conductivity is .ltoreq.about 10 mS/cm. 9. The method claim 1, wherein the cation exchange membrane is overloaded to a load density of 200-400 g/L. 10. The method of claim 1 wherein the cation exchange membrane has a pore size of 0.1 to 100 .mu.m. 11. The method of claim 1 wherein the cation exchange membrane is a cation exchange monolith or depth filter. 12. The method of claim 1, wherein the ionic polymer is polyethyleneimine (PEI). 13. The method of claim 1, wherein the antibody is a monoclonal antibody. 14. The method of claim 1, wherein the purification steps run continuously during steps a through e. 15. The method of claim 1, wherein the % yield of the antibody of interest is about 94%. 16. The method of claim 1, wherein the antibody of interest is trastuzumab. 17. The method of claim 1, wherein the antibody of interest is bevacizumab. 18. The method of claim 1, wherein the antibody of interest is rituximab. 19. The method of claim 1, wherein the antibody of interest is ranibizumab. 20. The method of any one of claims 1 and 16-19, further comprising preparing a pharmaceutical composition by combining the purified antibody with a pharmaceutically acceptable carrier. 21. The method of claim 1, wherein the dynamic binding capacity of the cation exchange chromatography purification step is improved at least 20-30%.

Details for Patent 10,364,268

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Genentech, Inc.	RITUXAN	rituximab	Injection	103705	11/26/1997		2039-08-19
Genentech, Inc.	HERCEPTIN	trastuzumab	For Injection	103792	09/25/1998		2039-08-19
Genentech, Inc.	HERCEPTIN	trastuzumab	For Injection	103792	02/10/2017		2039-08-19
Genentech, Inc.	AVASTIN	bevacizumab	Injection	125085	02/26/2004		2039-08-19