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用于蛋白质色谱的高渗透性和高容量整体柱的制备

Fabrication of high-permeability and high-capacity monolith for protein chromatography.

作者信息

Du Kai-Feng, Yang Dong, Sun Yan

机构信息

Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China.

出版信息

J Chromatogr A. 2007 Sep 7;1163(1-2):212-8. doi: 10.1016/j.chroma.2007.06.030. Epub 2007 Jun 26.

Abstract

A novel approach for the fabrication of macroporous poly(glycidyl methacrylate-ethylene glycol dimethacrylate) monolith is presented. The method involved the use of sodium sulfate granules and organic solvents as co-porogens. Compared with the conventional monoliths [ML-(1-3)] using organic solvents only as a porogen, the improved monoliths [MLS-(1-3)] showed not only higher column efficiency and dynamic binding capacity (DBC) for protein (bovine serum albumin, BSA), but also higher column permeability and lower back pressure. It is considered that the superpores introduced by the solid granules played an important role for the improvement of the monolith performance. Moreover, poly(glycidyl methacrylate-diethylamine) tentacles were grafted onto the pore surface of MLS-3 monolith. This has further increased the DBC of BSA to 74.7 mg/ml, about three times higher than that of the monoliths without the grafted tentacles. This grafting does not obviously decrease the column permeability, so a new monolith of high column permeability and binding capacity has been produced for high-performance preparative protein chromatography.

摘要

本文提出了一种制备大孔聚(甲基丙烯酸缩水甘油酯 - 乙二醇二甲基丙烯酸酯)整体柱的新方法。该方法使用硫酸钠颗粒和有机溶剂作为共致孔剂。与仅使用有机溶剂作为致孔剂的传统整体柱[ML-(1 - 3)]相比,改进后的整体柱[MLS-(1 - 3)]不仅对蛋白质(牛血清白蛋白,BSA)具有更高的柱效和动态结合容量(DBC),而且具有更高的柱渗透性和更低的背压。据认为,固体颗粒引入的超大孔对整体柱性能的改善起到了重要作用。此外,聚(甲基丙烯酸缩水甘油酯 - 二乙胺)触角被接枝到MLS-3整体柱的孔表面。这进一步将BSA的DBC提高到74.7 mg/ml,比没有接枝触角的整体柱高出约三倍。这种接枝不会明显降低柱渗透性,因此制备出了一种用于高效制备蛋白质色谱的具有高柱渗透性和结合容量的新型整体柱。

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