多功能环氧基填料固定蛋白A

摘要

The new multifunctional amino and epoxy support was used to immobilize proteins by two steps:First of all,the protein is physically absorbed by the amino group on the support;Then,the epoxy groups on the support covalently react with amino group of the adsorbed proteins to immobilized the pro -teins.Using the amino and epoxy support,the physical adsorption occurs immediately,and the concentra-tion of the supernatant of the immobilization suspension can be reduced to half within 10 min.Optimizing the condition of coupling protein A,and the optimal pH is 8.The optimum concentration of buffer solution is 5 mmol/L.As the ionic strength enhanced the immobilization capacity decreased.The ratio of greatest quality of amino group and epoxy group is 1:5(epoxy density is 75 μmol/g).The coupled efficiency is 90%in same condition above.The highest static and dynamic adsorption capacities of hIgG by this approach reached 38.9,31.4 mg/g,respectively.The amino and epoxy support has potential for the prep-aration of immune adsorption materials.%采用新的多功能氨基环氧基填料通过两步法固定蛋白A:①填料上的氨基物理吸附蛋白;②填料上的环氧基与蛋白上的氨基共价反应.此氨基环氧基填料对重组蛋白A的物理吸附发生的非常快,偶联时上清液浓度在10 min内立刻降至一半.优化了此填料与蛋白 A 偶联的条件.结果表明:最适 pH 为8,最佳盐溶液浓度为5 mmol/L,氨基与环氧基的最佳比为1:5(环氧基为75 μmol/g),在此条件下偶联率可达90%,用此方法合成的吸附材料对IgG的静态和动态吸附量最大值分别为38.9,31.4 mg/g.该氨基环氧基琼脂填料在制备免疫吸附材料领域具有良好的应用前景.