再生纤维素基固定化铜离子亲和膜上蛋白质吸附的分析

Analysis of protein adsorption on regenerated cellulose-based immobilized copper ion affinity membranes.

作者信息

Wu Chun-Yi, Suen Shing-Yi, Chen Shiow-Ching, Tzeng Jau-Hwan

机构信息

Department of Chemical Engineering, National Chung Hsing University, Taichung 402, Taiwan.

出版信息

J Chromatogr A. 2003 May 9;996(1-2):53-70. doi: 10.1016/s0021-9673(03)00531-4.

DOI:10.1016/s0021-9673(03)00531-4

PMID:12830908

Abstract

Immobilized metal affinity membranes (IMAMs) were prepared by immobilizing copper ions on microporous regenerated cellulose membranes through different types of chelating agents (dentate and triazine dye). The resulting chelator utilization percentages were 95% for iminodiacetic acid, 56% for N,N,N-tris(carboxymethyl)ethylenediamine, 52% for Cibacron blue 3GA, and 140% for Cibacron red 3BA. On the other hand, triazine dyes were slightly superior to dentate chelators on metal ion utilization for protein adsorption. In batch single-protein adsorptions, the protein adsorption capacity decreased with increasing molecular size and number of accessible surface histidine residues [lysozyme>bovine serum albumin(BSA)>gamma-globulin], while the binding strength order was the opposite (gamma-globulin>BSA>lysozyme). Moreover, the proportions of specific and nonspecific bindings were evaluated by varying pH and salt concentration conditions. A large fraction of the adsorption capacity was found to come from the nonspecific interactions for the prepared IMAMs. Lastly, batch three-protein adsorptions were performed and weak adsorption competition was observed.

摘要

通过不同类型的螯合剂（齿状螯合剂和三嗪染料）将铜离子固定在微孔再生纤维素膜上，制备了固定化金属亲和膜（IMAM）。生成的螯合剂利用率分别为：亚氨基二乙酸95%、N,N,N-三（羧甲基）乙二胺56%、汽巴克隆蓝3GA 52%、汽巴克隆红3BA 140%。另一方面，在用于蛋白质吸附的金属离子利用率方面，三嗪染料略优于齿状螯合剂。在分批单蛋白吸附中，蛋白质吸附容量随着分子大小和可及表面组氨酸残基数量的增加而降低[溶菌酶>牛血清白蛋白（BSA）>γ-球蛋白]，而结合强度顺序相反（γ-球蛋白>BSA>溶菌酶）。此外，通过改变pH和盐浓度条件评估了特异性和非特异性结合的比例。发现制备的IMAM的大部分吸附容量来自非特异性相互作用。最后，进行了分批三蛋白吸附，观察到弱吸附竞争。