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新型降钙素 (一种人降钙素类似物 )基因工程菌nCT/ pGEX 2T/E .coliBL2 1经 2 0L规模发酵 ,离心收集细胞 ,反复冻融裂解细胞 ,裂解液经G Sepharose 4B亲和层析纯化得到融合蛋白。融合蛋白进行磺酸化和溴化氰裂解后 ,用S Sepharose纯化得到新型降钙素前体 ,其纯度为 97 6 %。选用羧肽酶Y(CPY)将其前体转化为C 末端酰胺化的新型降钙素 ,转化率为 34 %。转化液经HPLC分离纯化得到新型降钙素。大鼠体内降血钙试验表明新型降钙素具有显著降血钙活性。
The new type of calcitonin (a human calcitonin analogue) genetically engineered bacteria nCT / pGEX 2T / E.coliBL2 1 by 20L scale fermentation, the cells were collected by centrifugation, repeatedly lysed and lysed cells, lysate by G Sepharose 4B affinity Chromatography purification to obtain the fusion protein. After sulfonation and cyanogen bromide cleavage of the fusion protein, a novel procalcitonin precursor was purified by S Sepharose with a purity of 97.6%. Carboxypeptidase Y (CPY) was used to convert its precursor to a C-terminal amidated novel calcitonin with a conversion of 34%. The conversion solution was separated and purified by HPLC to obtain new calcitonin. Hypocalcemia in rats showed that calcitonin has a significant hypocalcemic activity.