Proteomic Preparation Techniques for Formalin-Fixed Paraffin-Embedded Tissue Samples
10.19756/j.issn.0253-3820.241053
- VernacularTitle: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
- Author:
Ao LU
1
;
Bo MENG
;
Jia-Wei ZHAO
;
Huan-Yue LIAO
;
Zi-Hong YE
;
Xiang FANG
;
Yang ZHAO
Author Information
1. 中国计量大学生命科学学院,杭州 310018;中国计量科学研究院前沿计量科学中心,国家市场监管技术创新中心(质谱),北京 100029
- Keywords:
Formalin-fixed paraffin-embedded sample;
Sample preparation;
Proteomics;
Mass spectrometry;
Liquid chromatography-tandem mass spectroscopy
- From:
Chinese Journal of Analytical Chemistry
2025;53(1):84-93,中插4-中插8
- CountryChina
- Language:Chinese
-
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.
