Mobile phase effects on membrane protein elution during immobilized artificial membrane chromatography

Journal of Chromatography. a
C PidgeonC Bernal

Abstract

The eluotropic strength of different mobile phases for eluting membrane proteins from immobilized artificial membrane (IAM) chromatography surfaces was studied. Two protein mixtures containing bovine pancreatic PLA2 were used in this study. Protein mixture I was PLA2 obtained from Sigma which contained approximately 5-10 major protein bands in electrophoretic gels. Protein mixture II was obtained from flesh bovine pancreatic tissue and contained > 100 proteins including the target protein, PLA2. After adsorbing Sigma PLA2 to IAM columns, the elution conditions common to conventional chromatographic methods were evaluated for their ability to selectively purify PLA2. Elution conditions tested were (i) detergent gradients, (ii) salt gradients used during ion-exchange chromatography, (iii) salt conditions used during hydrophobic interaction chromatography, (iv) acetonitrile gradients used during reversed-phase chromatography, and (v) a two-step gradient consisting of first a detergent gradient followed by an acetonitrile gradient. Based on silver-stained electrophoretic protein gels. PLA2 from protein mixture I was purified to electrophoretic homogeneity with 417-fold increase in specific activity in one step using elution conditi...Continue Reading

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Citations

Jan 30, 2002·Biomedical Chromatography : BMC·Hiroshi Kamimori, Masaharu Konishi
Nov 6, 2001·Journal of Biochemical and Biophysical Methods·A Lundqvist, P Lundahl
Dec 16, 2000·Pharmaceutical Science & Technology Today·T Geetha, S Singh
Jul 17, 2007·Journal of Pharmaceutical Sciences·Ana Ruiz-GarciaVicente G Casabo
Oct 29, 2005·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Lutea A A de JongRainer Bischoff
Feb 3, 1998·Journal of Chromatography. B, Biomedical Sciences and Applications·H LiuC Pidgeon

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