Preparation of quaternary amine monolithic column for strong anion-exchange chromatography and its application to the separation of Enterovirus 71

被引:7
|
作者
Gu, Huimin [1 ]
Yin, Dezhong [1 ]
Ren, Jie [1 ]
Zhang, Baoliang [2 ]
Zhang, Qiuyu [2 ]
机构
[1] Northwestern Polytech Univ, Sch Sci, Xian 710072, Peoples R China
[2] Northwestern Polytech Univ, Key Lab Space Appl Phys & Chem, Minist Educ, Xian 710072, Peoples R China
基金
国家高技术研究发展计划(863计划);
关键词
Monolithic column; Strong anion-exchange chromatography; Enterovirus; 71; Quaternary amine; PERFORMANCE LIQUID-CHROMATOGRAPHY; MEMBRANE CHROMATOGRAPHY; MACROPOROUS POLYMER; STATIONARY PHASES; IMMUNE-RESPONSES; ION-EXCHANGE; PURIFICATION; EFFICIENT; VACCINES; MEDIA;
D O I
10.1016/j.jchromb.2016.09.020
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Large size virion is unable to diffuse into pores of conventional porous chromatography particles. Therefore, separation of virion by conventional column-packing materials is not quite efficient. To solve this problem, a monolithic column with large convective pores and quaternary amine groups was prepared and was applied to separate Enterovirus 71 (EV71, approximate to 5700-6000 kDa). Cross-section, pore structure, hydrodynamic performance, adsorption property and dynamic binding capacity of prepared monolithic column were determined. Double-pore structures, macropore at 2472 nm and mesopore at 5-60 nm, were formed. The porosity was up to 63.3%, which enable higher permeability and lower back pressure of the monolithic column than commercial UNO (TM) Q1 column. Based on the breakthrough curves, the loading capacity of bovine serum albumin was calculated to be 42.0 mg per column. In addition, prepared quaternary amine monolithic column was proved to be suitable for the separation of protein mixture by strong anion-exchange chromatography. As a practical application, prepared monolith column presents excellent performance to the separation of EV71 from virus-proteins mixture. (C) 2016 Elsevier B.V. All rights reserved.
引用
收藏
页码:399 / 405
页数:7
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