Single-step protein purification by back flush in ion exchange chromatography.
Ion exchange chromatography, one of the major procedures for protein purification, seldom provides single-step purification due to a lack of specific affinity. In this work, a novel and simple method called "back flush" (i.e., reversing the flow direction of elution relative to that of sample loading) was developed to achieve single-step purification on an ion exchanger. Tips for the conditions and operation by back flush are presented. Our study demonstrates, for the first time, the feasibility and dramatic improvement for protein purification by the back-flush method.
Chern
Ming-Kai
MK
Graduate Institute of Life Sciences, Tamkang University, Tamsui 25137, Taiwan. mkchern@mail.tku.edu.tw
Shiah
Wei-Jyh
WJ
Chen
Jyun-Jie
JJ
Tsai
Tzung-You
TY
Lin
Hsin-Yin
HY
Liu
Chien-Wei
CW
D000445:Aldehyde Oxidoreductases / Q000302:isolation & purification*; D002852:Chromatography, Ion Exchange / Q000295:instrumentation / Q000379:methods*; D020690:GTPase-Activating Proteins / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D011994:Recombinant Proteins / Q000302:isolation & purification; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D029701:Saccharomyces cerevisiae Proteins / Q000302:isolation & purification*
D020690:GTPase-Activating Proteins; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D000445:Aldehyde Oxidoreductases; C018699:aldehyde dehydrogenase (NAD(P)+)