上海医药2024,Vol.45Issue(15) :69-72.

蛋白提取工艺过程评价方法的研究

Study on the evaluation method for protein extraction process

梁吉 汪晨曦
上海医药2024,Vol.45Issue(15) :69-72.

蛋白提取工艺过程评价方法的研究

Study on the evaluation method for protein extraction process

梁吉 1汪晨曦2
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作者信息

  • 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
  • 折叠

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.

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,SDS-PAGE 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

Key words

SDS-PAGE/BCA/standard curve method/protein extraction/intermediate evaluation

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出版年

2024
上海医药
上海医药行业协会

上海医药

影响因子:0.781
ISSN:1006-1533
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