蛋白提取工艺过程评价方法的研究  

Study on the evaluation method for protein extraction process

作　　者：梁吉 汪晨曦 LIANG Ji;WANG Chenxi(Shanghai Pharmaceutical School,Shanghai 200135,China;SPH NO.1 Biochemical&Pharmaceutical Co.,Ltd.,Shanghai 200240,China)

机构地区：[1]上海市医药学校,上海200135 [2]上海上药第一生化药业有限公司,上海200240

出　　处：《上海医药》2024年第15期69-72,共4页Shanghai Medical & Pharmaceutical Journal

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。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,which provides a theoretical basis for subsequent process improvement.

关 键 词：SDS-PAGE BCA 标准曲线法 蛋白提取 中间体评价 

分 类 号：TQ460.63[化学工程—制药化工] TQ464.8