Effect of experimental conditions on strong biocomplimentary pairing in high-performance monolithic disk affinity chromatography

被引:37
作者
Ostryanina, ND [1 ]
Il'ina, OV [1 ]
Tennikova, TB [1 ]
机构
[1] Russian Acad Sci, Inst Macromol Cpds, St Petersburg 199004, Russia
来源
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES | 2002年 / 770卷 / 1-2期
关键词
biocomplenentary pairing; monolithic disks;
D O I
10.1016/S1570-0232(01)00597-9
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The effect of flow-rate on quantitatively determined binding parameters for several biocomplementary pairs in affinity mode high-performance monolithic disk affinity chromatography (HPMDAC) has been investigated using frontal analysis approach. Affinity interactions were evaluated from linearized adsorption isotherms and dynamic dissociation constants of the complexes K-diss, and the theoretical adsorption capacities Q(max) were calculated. HPMDAC isolation of atypical protein trypsin from both buffered solution and artificial mixture as well as biospecific extraction of antibodies against bovine serum albumin and recombinant protein G from such complex mixtures as blood serum and cellular lysate were examined. Immobilized counterparts soybean trypsin inhibitor, bovine serum albumin, and human immunoglobulin G were used in chromatographic experiments. The maximum adsorption capacities obtained at different flow-rates were compared with those determined at static conditions. The dependence of quantitative parameters on the surface density of immobilized ligands has also been explored. Finally, a series of experiments was carried out to evaluate the dependence of dynamic affinity binding on temperature for two complementary pairs. (C) 2002 Elsevier Science B V All rights reserved.
引用
收藏
页码:35 / 43
页数:9
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