江浙蝮蛇毒抗栓酶纯化方法的改进

Improvement of Purification Process of Antithrombotic Enzyme from Venin of Agkistrodon Halys Pallas

用DEAE－SephadexA50离子交换层析结合亲和层析方法，从江浙蝮蛇（AgkistrodonhalysPallas）粗毒中分离纯化抗栓酶（EC3．4．21．28）。所得抗栓酶在SDS－PAGE上呈一条主区带及二条副区带，分子量在33～44KD之间，比活为14000USP单位／mg蛋白，回收率可达到60％以上，制品的出血毒及神经毒素均可合格，较本所原制备方法的纯度和得率分别提高10倍和4倍，且操作简单，适于大规模制备。

The antithrombotic enzyme was purified from the crude venin of Agkistrodon halys pallas by DEAE-Sephadex A50 ion-exchange and affinity chromatography. On SDSPAGE profile, the purified antithrombotic enzyme showed a major band with a molecular weight of 33KD and two minor bands. The specific activity of the antithrombotic enzyme was 14000USP unit/mg protein, and the recovery rate of it was over 60%. Both the hemorrhagic toxicity and neurotoxicity of the enzyme were up to the specifications. Compared with those prepared with former process, the purity and recovery rate of the enzyme increased by 10-and 4-folds respectively. The improved process is easy to operate and suitable for large-scale production.