摘 要 目的：建立一种快速、便捷、低成本的生物酶制剂生产工艺过程评价方法。方法：联用凝胶电泳法和BCA蛋白定量法进行计算，得到较为准确的目标蛋白定量数量数据。结果：以胰蛋白酶生产工艺为例，对不同工段样品进行SDS-PAGE和BCA测试，并计算得到对应的蛋白定量数据，其中组内样品数据RSD≤5%，说明了该方法的可重复性。结论：该方法可准确分析不同工段的蛋白浓度变化，并进行快速、灵敏地定量；同时发现原胰蛋白酶生产工艺中存在蛋白泄漏的缺陷，可为后续工艺改进提供理论依据。

    关键词 SDS-PAGE BCA 标准曲线法 蛋白提取 中间体评价

    中图分类号：TQ460.63； TQ464.8 文献标志码：A 文章编号：1006-1533(2024)15-0069-04

    引用本文 梁吉， 汪晨曦. 蛋白提取工艺过程评价方法的研究[J]. 上海医药， 2024， 45(15)： 69-72.

    Study on the evaluation method for protein extraction process

    LIANG Ji1， WANG Chenxi2

    (1. Shanghai Pharmaceutical School， Shanghai 200135， China； 2. SPH NO.1 Biochemical & Pharmaceutical Co.， Ltd.， Shanghai 200240， China)

    ABSTRACT Objective： To establish a rapid， convenient and low-cost evaluation method for the production process of biological enzyme preparations. Methods： Combined with gel electrophoresis and BCA protein quantitative method， more accurate quantitative data of target protein were obtained. Results： Taking the trypsin production process as an example， SDSPAGE and BCA tests were performed on samples from different sections， and the corresponding protein quantitative data were obtained by calculation. The RSD≤5 % of the sample data in the group indicated the repeatability of the method. Conclusion： This method can accurately analyze the changes of protein concentration in different sections and quickly and sensitively quantify the protein. It is found that there is a defect of protein leakage in original production process of trypsin ......