多串心钠素的纯化与活性测定

为获得纯化心钠素(ANP)单体,采用离子交换及疏水柱层析,纯化融合蛋白麦芽糖结合蛋白(MBP)-ANP和MBP-3ANP,用凝血因子Xa切割MBP-ANP后,经阳离子柱分离获得ANP单体.对ANP单体与BMP-3ANP进行生物学活性检测.1　材料与方法1.1　材料含心钠素多拷贝基因的重组表达质粒pMal-nANP...

Purification of Multi-copied Atrial Natriuretic Peptide and the Identification of Its Activity

The expression product of α human atrial natriuretic peptide (ANP) was purified by ion exchange and hydrophobic chromatography. Purity of fusion protein maltose binding protein(MBP) ANP and MBP 3ANP was over 90%. After the MBP ANP was digested by factor Xa, the cation column was used to separate a single ANP. The purity of single ANP was 93%～96% by PAGE densitometric scan. The purified single ANP and MBP 3ANP caused dilation of previously contracted rat aorta slice successfully, which means it had good biological activity. Since the affinity resin is very expensive and its reproducibility is poor, it isn't suitable for scale up. Using ion exchange and hydrophobic chromatography instead of affinity chromatography, it showed that the separation of protein gave good result.