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Preparation of lysozyme molecularly imprinted polymers and purification of lysozyme from egg white

机译:溶菌酶分子印迹聚合物的制备及蛋清中溶菌酶的纯化

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摘要

Molecular imprinting as a promising and facile separation technique has received much attention because of its high selectivity for target molecules. In this study, lysozyme molecularly imprinted polymers (Lys-MIPs) were successfully prepared by the entrapment method with lysozyme as the template molecule, acrylamide as the functional monomer and N,N-methylenebisacrylamide as the cross-linker. The removal of the template lysozyme from the molecularly imprinted polymers was investigated in detail by two methods. The synthesized Lys-MIPs were characterized by scanning electron microscopy and Fourier transform-infrared, and the adsorption capacity, selectivity and reproducibility of the Lys-MIPs were also evaluated. The maximum adsorption capacity reached 94.8 mg/g, which is twice that of nonmolecularly imprinted polymers, and satisfactory selectivity and reproducibility were achieved. Using the Lys-MIP column, lysozyme could be separated completely from egg white, with purity close to 100% and mass recovery of 98.2%. This illustrated that the synthesized Lys-MIPs had high specific recognition and selectivity to the template lysozyme when they were applied to a mixture of protein standards and a real sample.
机译:分子印迹作为一种有前途且简便的分离技术,因其对靶分子的高选择性而备受关注。本研究以溶菌酶为模板分子,丙烯酰胺为功能单体,N,N-亚甲基双丙烯酰胺为交联剂,通过包埋法成功制备了溶菌酶分子印迹聚合物(Lys-MIPs)。通过两种方法详细研究了从分子印迹聚合物中去除模板溶菌酶的方法。通过扫描电子显微镜和傅立叶变换红外光谱对合成的Lys-MIPs进行表征,并评估了Lys-MIPs的吸附容量,选择性和重现性。最大吸附容量达到94.8 mg / g,是非分子印迹聚合物的两倍,并且具有令人满意的选择性和重现性。使用Lys-MIP柱,可以从蛋清中完全分离溶菌酶,纯度接近100%,质量回收率为98.2%。这说明当将合成的Lys-MIPs应用于蛋白质标准品和真实样品的混合物时,它们对模板溶菌酶具有很高的特异性识别和选择性。

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