Azimzadeh Omid, Atkinson Michael J, Tapio Soile
Institute of Radiation Biology, Helmholtz Zentrum München, German Research Center for Environmental Health, Ingolstaedter Landstrasse 1, Neuherberg, 85764, Germany.
Methods Mol Biol. 2015;1295:109-15. doi: 10.1007/978-1-4939-2550-6_10.
Formalin-fixed, paraffin-embedded (FFPE) tissue has recently gained interest as an alternative to fresh/frozen tissue for retrospective protein biomarker discovery. However, during the formalin fixation proteins undergo degradation and cross-linking, making conventional protein analysis technologies challenging. Cross-linking is even more challenging when quantitative proteome analysis of FFPE tissue is planned. The use of conventional protein labeling technologies on FFPE tissue has turned out to be problematic as the lysine residue labeling targets are frequently blocked by the formalin treatment. We have established a qualitative and quantitative proteomics analysis technique for FFPE tissues that combines label-free proteomic analysis with optimized protein extraction and separation conditions.
福尔马林固定、石蜡包埋(FFPE)组织最近作为新鲜/冷冻组织的替代物受到关注,用于回顾性蛋白质生物标志物的发现。然而,在福尔马林固定过程中,蛋白质会发生降解和交联,这使得传统的蛋白质分析技术面临挑战。当计划对FFPE组织进行定量蛋白质组分析时,交联带来的挑战更大。事实证明,在FFPE组织上使用传统的蛋白质标记技术存在问题,因为赖氨酸残基标记靶点经常被福尔马林处理所阻断。我们已经建立了一种针对FFPE组织的定性和定量蛋白质组学分析技术,该技术将无标记蛋白质组分析与优化的蛋白质提取和分离条件相结合。