固定化镍离子亲和层析胶的制备及其性能鉴定

Study on Preparation and Identification of Immobilized Metal Ion Affinity Adsorption Gel

以Sepharose 6B为原料 ,在强碱性条件下用环氧氯丙烷活化 ,再与亚氨基二乙酸 (IDA)的钠盐溶液反应 ,在活化好的胶颗粒表面接上很多手臂IDA ;最后与硫酸镍溶液反应 ,使手臂IDA与Ni2 +发生螯合反应 ,即得到固定化Ni2 +亲和层析胶 (Ni2 + IDA) .采用原子吸收法及从大肠杆菌表达产物中纯化重组人B淋巴细胞刺激因子 (hBLyS)等方法检测制备胶的理化指标和纯化蛋白质的性能 .结果表明用此法制得的亲和胶与相应商品胶的性能完全一致 ,而成本却不到商品胶的十分之一 .

Sepharose 6B was activated by epichlorohydrin in the strong base condition, and then reacted with solution of iminodiacetic sodium. The arms of IDA were conjuncted to the activated Sepharose 6B. Then the products were reacted with the solution of NiSO4. The arms of IDA were chelated with Ni2+, and the chelating resin Ni2+-IDA could be prepared. The physicochemical indexes and performance in purifying protein of the expressing product were assayed with atomic absorption method and purifying aimed protein-human B lymphocyte stimulator (hBLyS) from the expressing products in E. coli. The results indicated that the performance of made gel is very good, and its price is less than 1/10 of that of commodity gel.