Fast cation-exchange separation of proteins in a plastic microcapillary disc.
Fast cation-exchange separation of proteins in a plastic microcapillary disc.
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DOI：
10.1016/j.chroma.2011.01.030
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发表时间：
2011-03
期刊：
影响因子：
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通讯作者：
N. Darton;Nuno M. Reis;M. Mackley;Nigel K.H. Slater
中科院分区：
文献类型：
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作者：
N. Darton;Nuno M. Reis;M. Mackley;Nigel K.H. Slater
基于塑料微毛细管膜，开发了一种新型一次性蛋白质阳离子交换快速分离吸附材料。采用熔融挤出法从乙烯-乙烯醇共聚物（EVOH）制备了含有19个平行微毛细管的MCF，每个微毛细管的平均内径为142μm。将MCF表面官能化以产生阳离子交换吸附剂（本文称为MCF-EV 0 H-SP）。新MCF-EVOH-SP材料的动态结合能力通过前沿分析在标准液相色谱仪中使用纯蛋白质溶液在表观流速范围uLS=5.5-27.7cms-1下进行实验测定。对于一个5米长的薄膜，鸡蛋溶菌酶的平均动态结合容量约为100μg，配体结合密度为413 ngcm −2。动态结合能力在uLStested的范围内没有显著变化。这种新材料的应用，以阴离子BSA和阳离子溶菌酶在pH 7.2的差减色谱证明。两种阳离子蛋白质，溶菌酶和细胞色素-C，也进行了色谱分离，以期将这种技术应用于蛋白质的分析或纯化。未来的应用可能包括基于阴离子交换和其他吸附模式的分离。
A novel disposable adsorbent material for fast cation-exchange separation of proteins was developed based on plastic microcapillary films (MCFs). A MCF containing 19 parallel microcapillaries, each with a mean internal diameter of 142μm, was prepared using a melt extrusion process from an ethylene-vinyl alcohol copolymer (EVOH). The MCF was surface functionalised to produce a cation-exchange adsorbent (herein referred as MCF-EVOH-SP). The dynamic binding capacity of the new MCF-EVOH-SP material was experimentally determined by frontal analysis using pure protein solutions in a standard liquid chromatography instrument for a range of superficial flow velocities, uLS=5.5–27.7cms−1. The mean dynamic binding capacity for hen-egg lysozyme was found to be approximately 100μg for a 5m length film, giving a ligand binding density of 413ngcm−2. The dynamic binding capacity did not vary significantly over the range of uLStested. The application of this novel material to subtractive chromatography was demonstrated for anionic BSA and cationic lysozyme at pH 7.2. The chromatographic separation of two cationic proteins, lysozyme and cytochrome-c, was also performed with a view to applying this technology to the analysis or purification of proteins. Future applications might include separation based on anion exchange and other modes of adsorption.