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

被引:29
作者
Dubinina, NI
Kurenbin, OI
Tennikova, TB
机构
[1] Institute of Macromolecular Compounds, Russian Academy of Sciences
关键词
gradient elution; column length; proteins;
D O I
10.1016/S0021-9673(96)00545-6
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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 (X(0)) or the part of a column in which the quasi-steady state is attained was studied. The equation for estimating the X(0) 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.
引用
收藏
页码:217 / 225
页数:9
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