Ünlü Nuri, Ceylan Şeyda, Erzengin Mahmut, Odabaşı Mehmet
Aksaray University, Faculty of Arts and Science, Chemistry Department, Aksaray, Turkey.
J Sep Sci. 2011 Aug;34(16-17):2173-80. doi: 10.1002/jssc.201100269. Epub 2011 Jul 8.
As a low-cost natural adsorbent, diatomite (DA) (2 μm) has several advantages including high surface area, chemical reactivity, hydrophilicity and lack of toxicity. In this study, the protein adsorption performance of supermacroporous composite cryogels embedded with Ni(2+)-attached DA particles (Ni(2+)-ADAPs) was investigated. Supermacroporous poly(2-hydroxyethyl methacrylate) (PHEMA)-based monolithic composite cryogel column embedded with Ni(2+)-ADAPs was prepared by radical cryo-copolymerization of 2-hydroxyethyl methacrylate (HEMA) with N,N'-methylene-bis-acrylamide (MBAAm) as cross-linker directly in a plastic syringe for affinity purification of human serum albumin (HSA) both from aqueous solutions and human serum. The chemical composition and surface area of DA was determined by XRF and BET method, respectively. The characterization of composite cryogel was investigated by SEM. The effect of pH, and embedded Ni(2+)-ADAPs amount, initial HSA concentration, temperature and flow rate on adsorption were studied. The maximum amount of HSA adsorption from aqueous solution at pH 8.0 phosphate buffer was very high (485.15 mg/g DA). It was observed that HSA could be repeatedly adsorbed and desorbed to the embedded Ni(2+)-ADAPs in poly(2-hydroxyethyl methacrylate) composite cryogel without significant loss of adsorption capacity. The efficiency of albumin adsorption from human serum before and after albumin adsorption was also investigated with SDS-PAGE analyses.
作为一种低成本的天然吸附剂，硅藻土（DA）（2μm）具有诸多优点，包括高比表面积、化学反应性、亲水性和无毒性。在本研究中，对嵌入附着有Ni(2+)的硅藻土颗粒（Ni(2+)-ADAPs）的超大孔复合冷冻凝胶的蛋白质吸附性能进行了研究。通过甲基丙烯酸2-羟乙酯（HEMA）与N,N'-亚甲基双丙烯酰胺（MBAAm）作为交联剂在塑料注射器中直接进行自由基冷冻共聚，制备了嵌入Ni(2+)-ADAPs的基于超大孔聚甲基丙烯酸2-羟乙酯（PHEMA）的整体复合冷冻凝胶柱，用于从水溶液和人血清中亲和纯化人血清白蛋白（HSA）。分别通过XRF和BET方法测定了DA的化学成分和比表面积。通过扫描电子显微镜（SEM）对复合冷冻凝胶进行了表征。研究了pH、嵌入的Ni(2+)-ADAPs量、初始HSA浓度、温度和流速对吸附的影响。在pH 8.0的磷酸盐缓冲液中，从水溶液中吸附HSA的最大量非常高（485.15mg/g DA）。观察到HSA可以在聚甲基丙烯酸-2-羟乙酯复合冷冻凝胶中对嵌入的Ni(2+)-ADAPs进行反复吸附和解吸，而吸附容量没有明显损失。还通过SDS-PAGE分析研究了人血清在白蛋白吸附前后白蛋白的吸附效率。
