US 9,810,670 B2
Ionic strength-mediated pH gradient ion exchange chromatography
Yajun Wang, Foster City, CA (US); George Tony Moreno, South San Francisco, CA (US); Boyan Zhang, South San Francisco, CA (US); Liangyi Zhang, South San Francisco, CA (US); Dell Farnan, South San Francisco, CA (US); and Thomas Patapoff, South San Francisco, CA (US)
Assigned to GENENTECH, INC., South San Francisco, CA (US)
Appl. No. 14/443,329
Filed by Genentech, Inc., South San Francisco, CA (US)
PCT Filed Nov. 15, 2013, PCT No. PCT/US2013/070415
§ 371(c)(1), (2) Date May 15, 2015,
PCT Pub. No. WO2014/078729, PCT Pub. Date May 22, 2014.
Claims priority of provisional application 61/727,051, filed on Nov. 15, 2012.
Claims priority of provisional application 61/780,707, filed on Mar. 13, 2013.
Prior Publication US 2015/0285771 A1, Oct. 8, 2015
Int. Cl. G01N 30/96 (2006.01); C07K 1/18 (2006.01); G01N 33/68 (2006.01)
CPC G01N 30/96 (2013.01) [C07K 1/18 (2013.01); G01N 33/6803 (2013.01)] 46 Claims
 
1. A method for analyzing a composition comprising a polypeptide and one or more contaminants, the method comprising
a) binding the polypeptide and one of more contaminants to an ion-exchange chromatography material using a loading buffer, wherein the loading buffer is at an initial pH and comprises an initial ionic strength;
b) eluting the polypeptide and one or more contaminants from the ion-exchange chromatography material using an elution buffer wherein the pH of the elution buffer is altered in a pH gradient and wherein the ionic strength of the elution buffer is essentially the same as the initial ionic strength of the loading buffer, wherein the polypeptide and the one or more contaminants are separated by pH gradient at about the initial ionic strength; and
c) detecting the polypeptide and the one or more contaminants.