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环糊精在蛋白质毛细管区带电泳中作为选择性增强剂

Cyclodextrins as selectivity enhancers in capillary zone electrophoresis of proteins.

作者信息

Rathore A S, Horváth C

机构信息

Department of Chemical Engineering, Yale University New Haven, CT 06520-8286, USA.

出版信息

Electrophoresis. 1998 Oct;19(13):2285-9. doi: 10.1002/elps.1150191306.

Abstract

The selectivity in the capillary zone electrophoresis (CZE) of a variety of acidic and basic proteins including alpha-chymotrypsinogen A, cytochrome c, lysozyme, ribonuclease A, ovalbumin, and beta-lactoglobulins A and B, was altered by adding 6-monodeoxy-6-monoamino-beta-cyclodextrin or carboxymethylated beta-cyclodextrin to the electrophoretic medium of aqueous 50 mM sodium phosphate, pH 2.5. On the other hand, no significant improvement was obtained in the separation upon addition of heptakis (2,6-di-O-methyl)-beta-cyclodextrin. Whereas protein adsorption on the wall of raw silica capillaries was significant in the absence of cyclodextrin, by addition of beta-cyclodextrin or its derivatives to the background electrolyte, wall adsorption was reduced with concomitant enhancement of the recovery. The results confirm that in various separation techniques, particularly those which employ microcolumns, certain cyclodextrin additives can be useful selectivity enhancers not only in the separation of small sample molecules but also in that of proteins.

摘要

在50 mM磷酸钠水溶液(pH 2.5)的电泳介质中添加6-单脱氧-6-单氨基-β-环糊精或羧甲基化β-环糊精后,多种酸性和碱性蛋白质(包括α-胰凝乳蛋白酶原A、细胞色素c、溶菌酶、核糖核酸酶A、卵清蛋白以及β-乳球蛋白A和B)在毛细管区带电泳(CZE)中的选择性发生了改变。另一方面,添加七(2,6-二-O-甲基)-β-环糊精后,分离效果未得到显著改善。在没有环糊精的情况下,蛋白质在未处理的硅胶毛细管内壁上的吸附很明显,而向背景电解质中添加β-环糊精或其衍生物后,内壁吸附减少,回收率随之提高。结果证实,在各种分离技术中,尤其是那些使用微柱的技术,某些环糊精添加剂不仅在分离小样品分子方面,而且在蛋白质分离方面,都可能是有用的选择性增强剂。

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