分析化学2025，Vol.53Issue(1)：84-93,中插4-中插8,15.DOI:10.19756/j.issn.0253-3820.241053
福尔马林固定石蜡包埋组织样本的蛋白质组制备技术研究
Proteomic Preparation Techniques for Formalin-Fixed Paraffin-Embedded Tissue Samples
摘要
Abstract
Twelve pre-processing protocols for formalin-fixed paraffin-embedded(FFPE)tissue samples were developed by orthogonal experimental design,incorporating different dewaxing buffers(Triton X-100 and xylene),lysis buffers(TFE and RapiGest),and enzyme digestion methods(iST,SP3,and FASP)to explore the optimal experimental conditions.These protocols were assessed based on protein and peptide identification depth,identification stability,and quantitative levels of protein abundance.The results indicated that Triton X-100 and xylene minimally impacted proteomics identification,whereas the TFE lysis buffer and iST digestion method significantly enhanced the proteomics analysis of FFPE samples.Considering the potential toxicity of xylene,the TTI protocol based on Triton X-100,TFE,and iST was determined to be the optimal choice.This protocol exhibited the best repeatability and stability,and a higher number of proteins associated with significant biological functions were identified.In conclusion,the established TTI protocol offered an efficient and comprehensive approach for proteomic analysis of FFPE samples,significantly enhancing the repeatability and stability of protein identification.关键词
福尔马林固定石蜡包埋样本/样本制备/蛋白质组/质谱/液相色谱-串联质谱联用Key words
Formalin-fixed paraffin-embedded sample/Sample preparation/Proteomics/Mass spectrometry/Liquid chromatography-tandem mass spectroscopy引用本文复制引用
陆澳,孟波,赵佳威,廖焕玥,叶子弘,方向,赵洋..福尔马林固定石蜡包埋组织样本的蛋白质组制备技术研究[J].分析化学,2025,53(1):84-93,中插4-中插8,15.基金项目
科技基础资源调查专项项目(No.2022FY101202)、国家重点研发计划项目(No.2022YFF0608404)、国家自然科学基金项目(No.21927812)和中国计量科学研究院基本业务费重点项目(No.AKYZD2111)资助. Supported by the Science and Technology Basic Resources Survey Special Project(No.2022FY101202),the National Key Research and Development Program of China(No.2022YFF0608404),the National Natural Science Foundation of China(No.21927812)and the Research Project of the National Institute of Metrology(No.AKYZD2111). （No.2022FY101202）
