重组人表皮生长因子体外生物活性的测定  

Bioactivity Assay of Recombinant Human Epidermal Growth Factor

作　　者：何海琼[1] 王金发[1] 陈妍[1] 李黄金[2] 涂桂洪 

机构地区：[1]中山大学生命科学学院生物系,510275 [2]广州华生基因工程有限公司,510275

出　　处：《广东医学》1998年第5期324-325,共2页Guangdong Medical Journal

摘　　要：目的：探讨适合本实验室用M7TT或MTS／PMS检测重组人表皮生长因子（rhEGF）的一系列实验参数，包括血清浓度、细胞浓度、细胞培养时间和酶解程度。方法：利用MTF或MTS／PMS结合酶标仪分别在多种实验条件下对重组人表皮细胞因子进行了生物活性检测，同时绘制细胞生长曲线，对细胞生长状态拍照观察。结果：血清浓度为0．5％，细胞浓度为5000－7000个／孔，细胞铺满率为70％－80％及酶解适当时，用MTT和MTS／PMS都测得ED50约为0．6ng／ml，与标准品相符。结论：改进后的实验条件下，MTT和MTS／RMS都可用于rhEGF的生物活性检测。Objective:To determine the applicable conditions, including the concentration of serum, the number and the culturing period of cells and the extent of digestion or trypsin which have great effects on the bioactivity assay of EGF using MTT or MTS/PMS respectively. Methods: the bioectivity of rhEGF was assayed under kinds of conditions using MTT and MTS/PMS with ELISA. Result:under the condition of 0.5% serum, 5 000~7 000 cells/well, 70% ~ 80% spreading out and digested perfectly, the value of ED50 was determined as 0. 6 ng/ml. Conclusion: the modified conditions is appicable as to bioactivity assay with MTT or MTS/PMS in our lab.

关 键 词：表皮生长因子 重组 生物活性 

分 类 号：Q516[生物学—生物化学] TQ460.72[医药卫生—药物分析学]