title: Single-step protein purification by back flush in ion exchange chromatography.
abstract: 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.
authors.affiliation: Graduate Institute of Life Sciences, Tamkang University, Tamsui 25137, Taiwan. mkchern@mail.tku.edu.tw
authors.forename: Ming-Kai
authors.initials: MK
authors.lastname: Chern
authors.forename: Wei-Jyh
authors.initials: WJ
authors.lastname: Shiah
authors.forename: Jyun-Jie
authors.initials: JJ
authors.lastname: Chen
authors.forename: Tzung-You
authors.initials: TY
authors.lastname: Tsai
authors.forename: Hsin-Yin
authors.initials: HY
authors.lastname: Lin
authors.forename: Chien-Wei
authors.initials: CW
authors.lastname: Liu
mesh_terms: 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*
chemical_list: D020690:GTPase-Activating Proteins; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D000445:Aldehyde Oxidoreductases; C018699:aldehyde dehydrogenase (NAD(P)+)