Peculiarities of gradient ion-exchange high-performance liquid chromatography of proteins

Journal of Chromatography. a
N I DubininaT B Tennikova

Abstract

Since the influence of column length on protein resolution in high-performance liquid chromatography (HPLC) is not clear, different viewpoints presented in the literature are analysed in detail. The influence of gradient steepness on the length of the working column part (X0) or the part of a column in which the quasi-steady state is attained was studied. The equation for estimating the X0 value was obtained for the general case of the retention model. It was shown that at steep gradients only a short part of the column is used as the working part on which all separation processes develop. The other part of a column is a ballast where the protein zone migrates in a regime of parallel transfer. These results form a theoretical basis for high-performance membrane chromatography. As was shown experimentally, this method makes it possible to perform protein separation at low gradient times with appropriate resolution, comparable with that of HPLC.

Citations

Mar 8, 2013·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Aleš PodgornikNika Lendero Krajnc
Sep 10, 2004·Journal of Separation Science·Alfred ZöchlingAlois Jungbauer
Aug 12, 2009·Journal of Separation Science·Ngoc Phuoc DinhKnut Irgum
Jan 8, 2013·Journal of Separation Science·Evgenia G Vlakh, Tatiana B Tennikova
Sep 4, 2004·Journal of Biochemical and Biophysical Methods·A PodgornikA Strancar
Mar 3, 2011·Journal of Biomaterials Applications·J Vincent EdwardsSteven C Goheen
Mar 24, 2005·Journal of Chromatography. a·Tatiana B Tennikova, Joachim Reusch
Oct 31, 2000·Analytical Chemistry·R Hahn, A Jungbauer
Sep 3, 1998·Analytical Chemistry·R GiovanniniT B Tennikova
Aug 1, 1999·Analytical Chemistry·A PodgornikT Tennikova

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