Clinical proteomics: Current status, challenges, and future perspectives

This account will give an overview and evaluation of the current advances in mass spectrometry (MS)-based proteomics platforms and technology. A general review of some background information concerning the application of these methods in the characterization of molecular sizes and related protein expression profiles associated with different types of cells under varied experimental conditions will be presented. It is intended to provide a concise and succinct overview to those clinical researchers first exposed to this foremost powerful methodology in modern life sciences of postgenomic era. Proteomic characterization using highly sophisticated and expensive instrumentation of MS has been used to characterize biological samples of complex protein mixtures with vastly different protein structure and composition. These systems are then used to highlight the versatility and potential of the MS-based proteomic strategies for facilitating protein expression analysis of various disease-related organisms or tissues of interest. Major MS-based strategies reviewed herein include (1) matrix-assisted laser desorption ionization-MS and electron-spray ionization proteomics; (2) one-dimensional or two-dimensional gel-based proteomics; (3) gel-free shotgun proteomics in conjunction with liquid chromatography/tandem MS; (4) Multiple reaction monitoring coupled tandem MS quantitative proteomics and; (5) Phosphoproteomics based on immobilized metal affinity chromatography and liquid chromatography-MS/MS.

摘要本評論文章將針對以質譜為基礎的蛋白質體學平台和技術的進展做一現況回顧和評價。文章中將廣泛性回顧過去應用質譜方法,分析鑑定來自各種實驗條件下不同來源細胞的蛋白質分子大小與整體蛋白質表現圖譜。本文章的目的是針對首次接觸到這個最重要且功能強大的質譜學新技術方法的臨床研究人員,在後基因體時代應該具備的簡明基礎性蛋白質體學概念。運用高度複雜和昂貴質譜儀器的蛋白質體鑑定技術,已被用於鑑定具有截然不同的結構和組成的複雜蛋白質混合物生物樣品。這些顯著且具有多功能性和有潛力性的系統則已被有效應用於各種疾病相關的微生物或組織的蛋白質體表達分析。這些以質譜為基礎的主要策略回顧包括：1.基質輔助雷射脫附游離質譜和電灑游離的蛋白質體學 2.一維或二維凝膠電泳的蛋白質體學 3.無凝膠散彈槍法蛋白質體學連接液相層析/串聯質譜（LC-MS/MS）4.多重反應監控掃瞄耦合串聯質譜（MS/MS-MRM）的定量蛋白質體學 5.利用固定化金屬親和層析（IMAC）結合液相層析串聯質譜的磷酸化蛋白質體學。.