US 11,912,739 B2
Sterile chromatography and manufacturing processes
Rahul Godawat, Bridgewater, NJ (US); Veena Warikoo, Bridgewater, NJ (US); Rohan Patil, Bridgewater, NJ (US); Konstantin Konstantinov, Bridgewater, NJ (US); Venkat Kishore Ryakala, Bridgewater, NJ (US); and Mahsa Rohani, Bridgewater, NJ (US)
Assigned to Genzyme Corporation, Cambridge, MA (US)
Filed by Genzyme Corporation, Cambridge, MA (US)
Filed on Aug. 14, 2020, as Appl. No. 16/993,654.
Application 16/993,654 is a continuation of application No. 16/107,203, filed on Aug. 21, 2018, abandoned.
Application 16/107,203 is a continuation of application No. 14/598,450, filed on Jan. 16, 2015, granted, now 10,087,214, issued on Oct. 2, 2018.
Claims priority of provisional application 61/928,906, filed on Jan. 17, 2014.
Prior Publication US 2020/0369718 A1, Nov. 26, 2020
Int. Cl. C07K 1/16 (2006.01); A61L 2/08 (2006.01); B01D 15/18 (2006.01); B01D 15/20 (2006.01); B01D 15/36 (2006.01); B01D 15/38 (2006.01); B01D 15/42 (2006.01); C07K 1/22 (2006.01); C12N 9/40 (2006.01)
CPC C07K 1/16 (2013.01) [A61L 2/081 (2013.01); B01D 15/1864 (2013.01); B01D 15/20 (2013.01); B01D 15/363 (2013.01); B01D 15/3809 (2013.01); B01D 15/426 (2013.01); C07K 1/165 (2013.01); C07K 1/22 (2013.01); C12N 9/2465 (2013.01); B01D 15/1807 (2013.01); C12Y 302/01022 (2013.01)] 20 Claims
OG exemplary drawing
 
1. A method of performing chromatography with a gamma-irradiated chromatography resin, comprising:
(a) providing a single chromatography column containing a gamma-irradiated chromatography resin, wherein the gamma-irradiated chromatography resin has been treated with a dose of gamma-irradiation between about 10 kGy and about 40 kGy; and
(b) performing at least two cycles of chromatography with the single chromatography column, wherein:
each cycle of the at least two cycles of chromatography comprises the steps of:
capturing a recombinant protein by exposing the gamma-irradiated chromatography resin to a liquid containing the recombinant protein;
washing the gamma-irradiated chromatography resin by exposing the gamma-irradiated chromatography resin to a wash buffer;
eluting the recombinant protein by exposing the gamma-irradiated chromatography resin to an elution buffer; and
recovering binding capacity of the gamma-irradiated chromatography resin by exposing the gamma-irradiated chromatography resin to a denaturing buffer;
the denaturing buffer comprises one or more of urea, guanidine hydrochloride, 2-[4-(2,4,4,-trimethypentan-2-yl)phenoxy]ethanol, acetic acid, and arginine; and
during each of the at least two cycles the flow rate, volume, and concentration of the denaturing buffer are selected to recover binding capacity of the gamma-irradiated chromatography resin.