Immunoglobulin G purification from bovine serum with pseudo-specific supermacroporous cryogels
Immunoglobulin G purification from bovine serum with pseudo-specific supermacroporous cryogels
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DOI：
10.1016/j.seppur.2013.08.026
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发表时间：
2013-10-30
影响因子：
8.6
通讯作者：
Denizli, Adil
中科院分区：
文献类型：
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作者：
Daoud-Attieh, Mira;Chaib, Haidar;Denizli, Adil
合成了超大孔冷冻胶，并将其用于从牛血清中分离纯化免疫球蛋白G。以N-乙烯基咪唑为共聚单体，将咪唑官能团引入到碱性聚合物骨架中，获得了对酸凝胶的假特异性。在部分冷冻在塑料注射管柱中的单体的水溶液中，通过冷冻凝胶法制备了冷冻机。以水溶性功能单体n-乙烯基咪唑与碱性单体HEMA共聚，制备了聚(甲基丙烯酸羟乙酯-n-乙烯基咪唑)[聚(HEMA-VIM)]冷冻胶。采用溶胀试验、傅里叶变换红外光谱(FTIR)、元素分析和扫描电子显微镜(SEM)等表征手段对所制得的冷冻胶的理化性能进行了表征。研究了不同pH值、不同浓度、不同流速、不同离子强度、不同吸附时间等因素对牛免疫球蛋白在普通和复合冷冻机上的吸附效果。在pH为7.4~21.1 mg/单位质量的聚乙二醇凝胶中，牛免疫球蛋白的吸附效果最好。在pH为4.0的条件下，用含有I M NaCI的0.1M醋酸盐缓冲液可以很容易地洗脱吸附在凝胶中的抗体。为了描述吸附过程，我们用一些平衡和动力学吸附模型来描述数据。实验结果符合Langmuir模型，说明冷冻胶对蛋白质的单层吸附和表面均一性。最后，在确定的最佳条件下，对牛血清中的免疫球蛋白进行了纯化。用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法(SDS-PAGE)检测纯化效率和免疫球蛋白纯度。(C)2013爱思唯尔B.V.保留所有权利。
Supermacroporous cryogels were synthesized and used for the purification of immunoglobulin G (IgG) from bovine serum. Imidazole functional groups were incorporated into basic polymeric backbone to gain pseudo-specificity to ciyogels by using n-vinylimidazole as comonomer. The cryogels were prepared in aqueous solution of monomers partially frozen inside plastic syringe column by cryogelation. Poly (2-hydroxyethyl methacrylate-n-vinylimidazole) [poly(HEMA-VIM)] cryogels were prepared by polymerization of water-soluble functional monomer, n-vinylimidazole, with basic monomer HEMA. The characterization methods including swelling test, Fourier transform infrared spectroscopy (FTIR), elemental analysis, and scanning electron microscopy (SEM) were performed to evaluate physical and chemical properties of cryogels prepared. Bovine IgG adsorption on plain and composite cryogels was studied with respect to different parameters such as, pH, IgG concentration, flow rate, ionic strengths, and adsorption time. The best adsorption of bovine IgG was observed at pH 7.4 up to 21.1 mg per unit mass of poly (HEMA-VIM) ciyogel. Elution of IgG adsorbed from the ayogels was easily achieved with 0.1 M acetate buffer containing I M NaCI at pH 4.0. In order to describe the adsorption process, we applied some equilibrium and kinetic adsorption models to the data. The results best fitted to Langmuir model showing monolayer protein adsorption and surface homogeneity of cryogel. Finally, we evaluated IgG purification from bovine serum under optimal condition determined. The purification efficiency and IgG purity were investigated with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) study. (C) 2013 Elsevier B.V. All rights reserved.