一种制备poly(Ⅰ)·poly(C)-滤纸的新方法

poly(1)·poly(C)-滤纸是一种亲和材料,可以用来吸附与双链核酸有亲和力的酶或蛋白。本文介绍用对-β硫酸酯乙砜基苯胺为活化剂制备poly(I)·poly(C)-滤纸的方法。poly(I)·poly(C)的结合容量为10—35μg/cm~2,用来吸附兔网织红细胞裂解液中2’-5’A合成酶效果良好。在一定范围内,酶活与被吸附裂解液量呈线性关系,说明可以用来定量检测未知样品中与poly(I)·poly(C)有亲和力的酶。poly(I)·poly(C)-滤纸在-20℃保存四个月亲和能力不变。本方法与文献报道的方法相比,操作简便试剂易得。

A novel metnod for preparing poly(I).poly(C)-paper

Instead of l-[(m-nitrobenzyloxy) methyl]pyridinium chloride, β-sulfatoethyl s/&&&lphonyl aniline was used to activate Whatman 3 MM paper and aminobenzene sulphonylethyl paper was obtained. After diazotization poly(I). poly(C) was coupled on diazobenzenesulphonylethyl paper. The poly (I) .poly (C)-paper with capacity of 10-35>μg of poly(I) . poly(C)/cm2 could be used as affinity material to specifically adsorb 2'-5'A synthetase. Under optimal condition (3 mM ATP, 24-40 mM Mg (OAc)2, 30℃ for 18 hours) the bound enzyme catalyzed the conversion of more than 30% of ATP into 2'-5'A. The synthesis of 2'-5'A was linearly proportional to the amounts of rabbit reticulocyte lysate used in the assay ranging from 10ul up to 300ul. Compared with the method reported by G.R.Stark et al. this method offered some advantages, simple to prepare β-sulfatoethylsulphonyl aniline readily available and low cost.