红细胞嘧啶5′核苷酸酶的纯化及其抗血清的制备

Purification of Human Erythrocyte Pyrimidine-5′-Nucleotidase and Preparation of Rabbit Anibody Specific for Human P5′N.

利用UMP ADH Sepharose4B亲和层析的方法纯化正常人红细胞嘧啶 5′ 核苷酸酶 (P5′N ,EC 3 1 3 5) .结果表明 :利用UMP ADH Sepharose4B亲和层析柱可有效、专一性吸附P5′N ,纯化液经聚丙烯酰胺凝胶电泳显示一条蛋白质带 ,测定纯化物相对分子质量 2 8× 10 3 ,等电点 ( pI) 5 2 .纯化物免疫家兔后 ,得兔抗人P5′N抗血清 。

After the UMP ADH Sephsrose 4B column was made as the affinity manterial,human erythrocyte pyrimidine 5′ nucleotidase(P5′N EC 3 1 3 5 ) was purified from the blood of normal subjects by a combination of DEAE cellulose chromatography 、ammonium sulfate fractionation、ion exchange and affinity chromatography. The results show that the P5′N can adhere to the UMP ADH Sephsrose 4B column tightly with high specificity. Polyacrylamide electrophoresis of the purified material shows one strong protein band. The enzyme has p I of 5 2 and a relative molecular mass of 28 000 by polyacrylamide electrophoresis.The anyibody was obtained after the rabbits were immunized with the purified enzyme. It might play an important role in clinical study on hereditary or acquired erythrocyte pyrimidine 5′ nucleotidase dificiency.