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[新型共聚物涂层毛细管电泳柱及其在蛋白质分离中的应用]

[The novel copolymer coated capillary columns of electrophoresis and their applications to separation of proteins].

作者信息

Lu G, Gao D, Gu J, Fu R, Li F, Zhang H

机构信息

Department of Chemical Engineering & Material Sciences, Beijing Institute of Technology, Beijing, 100081.

出版信息

Se Pu. 1999 Jan;17(1):64-6.

Abstract

The copolymer of acrylonitrile, methyl acrylate, hydroxy ethyl acrylate (ZB-004), the copolymer of acrylonitrile, methyl acrylate, hydroxy ethyl acrylate, acrylamide (ZB-014) and the copolymer of acrylonitrile, hydroxy ethyl acrylate (ZB-016) were coated on the inner surface of fused-silica capillaries by just filling the capillary with solutions containing these copolymers followed by flushing the capillary with nitrogen. The physically adsorbed layer can reduce both protein adsorption and electroosmotic flow in the pH range of 3-5. Electroosmotic flow decreased by raising the concentrations of the copolymers. Separation performance of ZB-004 layer is better than those of other two layers due to its low hydrophilicity, but with higher pH values, appreciable peak deformation and increase in electroosmosis were observed. The intra day and inter day migration reproducibility were investigated in terms of relative standard deviation (RSD) with four basic proteins at pH 4.0. The RSDs of the intra day migration times were less than 2%. The RSDs of the inter day migration times were less than 4%. At pH 5.0, the RSDs of the migration times in two ZB-004-coated capillaries made on two different days were less than 1%. Separation efficiencies of four basic proteins in a ZB-004-coated capillary which stored in a buffer (pH 4.0) for fifteen days after being used for 14 days decreased 15%. These coatings were stable and exhibited reproducible separations from intra day, inter day and inter column under acidic conditions.

摘要

将丙烯腈、丙烯酸甲酯、丙烯酸羟乙酯的共聚物(ZB - 004)、丙烯腈、丙烯酸甲酯、丙烯酸羟乙酯、丙烯酰胺的共聚物(ZB - 014)以及丙烯腈、丙烯酸羟乙酯的共聚物(ZB - 016)涂覆在熔融石英毛细管的内表面,方法是将含有这些共聚物的溶液注入毛细管,然后用氮气冲洗毛细管。物理吸附层可在pH值为3 - 5的范围内减少蛋白质吸附和电渗流。通过提高共聚物的浓度,电渗流会降低。由于ZB - 004层的亲水性较低,其分离性能优于其他两层,但在较高pH值时,观察到明显的峰变形和电渗增加。在pH 4.0条件下,以四种碱性蛋白的相对标准偏差(RSD)研究了日内和日间迁移重现性。日内迁移时间的RSD小于2%。日间迁移时间的RSD小于4%。在pH 5.0时,两根在不同日期制备的涂有ZB - 004的毛细管中迁移时间的RSD小于1%。一根涂有ZB - 004的毛细管在使用14天后于缓冲液(pH 4.0)中储存15天,四种碱性蛋白的分离效率降低了15%。这些涂层稳定,在酸性条件下,日内、日间和柱间均表现出可重现的分离。

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