Immobilized metal affinity monolithic cryogels for cytochrome c purification
Immobilized metal affinity monolithic cryogels for cytochrome c purification
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DOI：
10.1016/j.colsurfb.2011.11.058
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发表时间：
2012-05-01
影响因子：
5.8
通讯作者：
Denizli, Adil
中科院分区：
文献类型：
--
作者：
Cimen, Duygu;Denizli, Adil
制备了一种高效、经济、快速纯化大鼠肝匀浆细胞色素c的冷冻凝胶整体柱。使用N-甲基丙烯酰基-（L）-组氨酸甲酯（MAR）作为金属螯合配体。以N，N，N '，N'-四亚甲基二胺（TEMED）和过硫酸铵（APS）为引发剂，在冰浴中通过自由基聚合制备了聚（甲基丙烯酸羟乙酯-N-甲基丙烯酰基-（L）-组氨酸甲酯）[PHEMAH]整体晶胶。通过比表面积测量、溶胀研究、扫描电子显微镜和元素分析对PHEMAH冻凝胶进行了表征。PHEMAH冻凝胶的比表面积为38.6m2/g。PHEMAH冻凝胶（MAH含量为113.7 μ mol/g）与Cu ~（2+）通过MAH直接络合，从水溶液和大鼠肝匀浆中吸附细胞色素c。细胞色素c在PHEMAH冻凝胶上的吸附量为3.1mg/g。发现在pH 8.0的磷酸盐缓冲液中，Cu 2+螯合的PHEMAH晶凝胶（携带58.7 μ mol Cu 2 +/g聚合物）的最大细胞色素c吸附容量为20.8 mg/g。大鼠肝匀浆中细胞色素c的吸附量为37.7mg/g，纯度为93.8%。观察到细胞色素c可以被PHEMAH冻凝胶反复吸附和洗脱，吸附能力没有明显损失。(C)2012爱思唯尔有限公司版权所有。
A monolithic cryogel column was prepared to obtain an efficient, cost effective and rapid purification of cytochrome c from rat liver homogenate. N-Methacryloyl-(L)-histidine methyl ester (MAR) was used as the metal-chelating ligand. Poly(hydroxyethyl methacrylate-N-methacryloyl-(L)-histidine methyl ester) [PHEMAH] monolithic cryogel was produced by free radical polymerization initiated by N,N,N',N'-tetramethylene diamine (TEMED) and ammonium persulfate (APS) pair in an ice bath. PHEMAH cryogel was characterized by surface area measurements, swelling studies, scanning electron microscopy and elemental analysis. The PHEMAH cryogel had a specific surface area of 38.6 m(2)/g. The PHEMAH cryogel containing 113.7 mu mol MAH/g was complexed with the Cu2+ ions directly via MAH for the adsorption of cytochrome c from aqueous solutions and rat liver homogenate. The cytochrome c adsorption on the PHEMAH cryogel was 3.1 mg/g. The maximum cytochrome c adsorption capacity of the Cu2+-chelated PHEMAH cryogel (carrying 58.7 mu mol Cu2+ per gram of polymer) was found to be 20.8 mg/g at pH 8.0 in phosphate buffer. The cytochrome c adsorption amount from rat liver homogenate was 37.7 mg/g with a purity of 93.8%. It was observed that cytochrome c could be repeatedly adsorbed and eluted with the PHEMAH cryogel without significant loss in the adsorption capacity. (C) 2012 Elsevier B.V. All rights reserved.