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 |
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.
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