通过免疫亲和色谱法从斑海豹（Phoca vitulina larga）肠道内容物中纯化碱性磷酸酶。

Purification of alkaline phosphatase from the intestinal content of common seal (Phoca vitulina larga) by immunoaffinity chromatography.

Laboratory of Biologically Active Substances of Hydrobionts, Ministry of Health, Moscow, USSR.

Comp Biochem Physiol B. 1991;99(3):509-11. doi: 10.1016/0305-0491(91)90330-g.

Abstract

A monoclonal antibody APP, 1 against harp seal alkaline phosphatase has been prepared. It was found that the antibody was cross-reacted with the intestinal alkaline phosphatase of common seal Phoca vitulina larga. 2. Purified antibody was linked to Sepharose 4B and used for immunoaffinity chromatographic purification of alkaline phosphatase from the intestinal content of common seal. A spec. act. of the purified enzyme was 7300 units per mg of protein. 3. The enzyme in 7.5% polyacrylamide gel in the presence of 2-mercaptoethanol and SDS was migrated as a single band of Mr 67,000. The value of the apparent Km for common seal alkaline phosphatase was equal to 3.7 mM.

摘要

制备了一种针对竖琴海豹碱性磷酸酶的单克隆抗体APP1。发现该抗体与斑海豹（Phoca vitulina larga）的肠碱性磷酸酶发生交叉反应。2. 将纯化的抗体与琼脂糖4B相连，用于从斑海豹的肠内容物中免疫亲和层析纯化碱性磷酸酶。纯化酶的比活性为每毫克蛋白质7300单位。3. 在2-巯基乙醇和十二烷基硫酸钠存在的情况下，该酶在7.5%聚丙烯酰胺凝胶中迁移为一条Mr 67,000的单一带。斑海豹碱性磷酸酶的表观Km值等于3.7 mM。