US 11,945,837 B2
Ion exchange membrane chromatography
Jerome Joseph Bill, Jr., South San Francisco, CA (US); Arick Michael Brown, South San Francisco, CA (US); Christopher John Dowd, South San Francisco, CA (US); and Brooke Ellen Thayer, South San Francisco, CA (US)
Assigned to Genentech, Inc., South San Francisco, CA (US)
Filed by Genentech, Inc., South San Francisco, CA (US)
Filed on Jun. 6, 2019, as Appl. No. 16/433,763.
Application 16/433,763 is a continuation of application No. 14/365,449, granted, now 10,364,268, previously published as PCT/US2012/070373, filed on Dec. 18, 2012.
Claims priority of provisional application 61/579,285, filed on Dec. 22, 2011.
Prior Publication US 2020/0102346 A1, Apr. 2, 2020
This patent is subject to a terminal disclaimer.
Int. Cl. C07K 1/18 (2006.01); B01D 15/36 (2006.01); C07K 1/16 (2006.01); C07K 16/06 (2006.01)
CPC C07K 1/18 (2013.01) [C07K 1/165 (2013.01); C07K 16/065 (2013.01)] 16 Claims
 
1. A method of enhancing efficiency of downstream chromatography steps for purification of proteins comprising:
a. passing a composition comprising a polypeptide of interest and one or more contaminants through a cation exchange membrane, wherein the one or more contaminants is one or more of a host cell protein, aminoglycoside antibiotic, nucleic acid, a variant of the polypeptide of interest, another polypeptide, an endotoxin, or a viral contaminant, 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 cation 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 cation exchange membrane comprising the polypeptide of interest;
d. subjecting the membrane effluent comprising the polypeptide of interest to a cation exchange chromatography step, and
e. recovering the purified polypeptide from the effluent of the cation exchange chromatography step.