生物学杂志

生物学杂志 ›› 2025, Vol. 42 ›› Issue (4): 38-.doi: 10.3969/j.issn.2095-1736.2025.04.038

• 研究报告 • 上一篇    下一篇

基于特征多肽的间充质干细胞裂解物质量控制方法研究

邢芳毓1,2， 徐丽明3， 张贵锋1,2   

  1. 1. 中国科学院过程工程研究所 生物药制备与递送全国重点实验室， 北京 100190；
    2. 中国科学院大学 化学工程学院， 北京 100049；
    3. 中国食品药品检定研究院， 北京 102629
  • 出版日期:2025-08-18 发布日期:2025-08-13
  • 通讯作者: 张贵锋，博士，研究员，研究方向为生物化工，E-mail:gfzhang@ipe.ac.cn
  • 作者简介:邢芳毓，博士研究生，研究方向为生物化工，E-mail:xingfangyu21@ipe.ac.cn
  • 基金资助:
    北京市自然科学基金-昌平创新联合基金项目（L234021， L244034）

Study on quality control of mesenchymal stem cell lysates using marker peptides

XING Fangyu1,2, XU Liming3, ZHANG Guifeng1,2   

  1. 1. State Key Laboratory of Biopharmaceutical Preparation and Delivery, Institute of Process Engineering, Chinese
    Academy of Sciences, Beijing 100190, China； 2. School of Chemical and Engineering, University of Chinese Academy
    of Sciences, Beijing 100049, China； 3. National Institutes for Food and Drug Control, Beijing 102629, China
  • Online:2025-08-18 Published:2025-08-13

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摘要： 利用生物质谱技术研究间充质干细胞裂解物中的蛋白质组成并尝试建立质量控制方法。首先利用液相色谱-质谱联用技术分析间充质干细胞裂解物中的蛋白质组成，筛选出高丰度蛋白质，利用特征多肽法建立高丰度蛋白质定量方法，并进行方法学考察。利用质谱技术从间充质干细胞裂解物中识别出369种蛋白质，从中筛选出3种高丰度蛋白（波形蛋白、细丝蛋白A和膜联蛋白A2）作为组成稳定性评价指标；利用特征多肽法实现3种高丰度蛋白质的定量检测；方法学实验结果表明，方法的添加回收率满足规定，方法重复性的相对标准偏差低于3.18%，且方法的检测下限为皮克级。实验结果表明，利用高丰度蛋白质作为指标评价间充质干细胞裂解物质量稳定性具有可行性。

关键词: 间充质干细胞裂解物, 特征多肽, 液相色谱-质谱联用技术, 高丰度蛋白质定量, 质量控制

Abstract: Using biomass spectrometry, the protein contents of the mesenchymal stem cell (MSC) lysates were studied, and a quality control method was developed. The protein composition of MSC lysates was analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS), then the high-abundance proteins were screened out. Method for the quantification of high-abundance proteins was established using the method of marker peptides, and a methodological investigation was carried out. Through LC-MS/MS analysis, 369 proteins were identified from MSC lysates, among which three high-abundance proteins (Vimentin, Filamin-A, and Annexin A2) were screened out as the evaluation indexes of compositional stability; the quantitative detection of the three high-abundance proteins was achieved by using the method of marker peptides; and the results of the methodological experiments showed that the spiked recoveries were satisfactory, the relative standard deviations of the method repeatability were lower than 3.18%, and the lower limit of detection of the method was at the pg level. The results demonstrated that it was feasible to assess the quality and stability of MSC lysates using high-abundance proteins as markers.

Key words: mesenchymal stem cell lysates, marker peptides, liquid chromatography-tandem mass spectrometry, high abundance protein quantification, quality contro

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