基于二维凝胶电泳的蛋白质定量分析技术

Quantitative Analytical Techniques of Proteins Based on Two-Dimensional Gel Electrophoresis

基于2DE的蛋白质定量分析技术通过对不同样品蛋白质表达差异的比较,定量地解析了蛋白质的动态变化,为了解和阐明细胞蛋白质功能及活动机制等提供了重要信息。本文简单介绍了传统2DE的“一个样品一块胶”模式的定量分析技术,指出其因为耗时、繁琐及重复性较差等,在常规2DE定量分析应用中受到了制约;阐述了近年来发展的“多样品共分离”模式的定量技术,包括荧光胶内差示电泳(DIGE)、“不同胶曝光”以及稳定同位素标记技术,这类技术因有效克服了传统技术的局限性,由此提高了定量分析的能力;分别对每种蛋白质定量技术及其优缺点作了比较和评述,并对2DE定量分析技术的未来发展方向做出了展望。

Quantitative analytical techniques of proteins based on two-dimensional gel electrophoresis, depending on the comparing of altered levels of proteins between different samples to obtain quantitative changes, are informative for comprehesive understanding functions and biological mechanism of proteins in cells. Conventional methods in which protein samples are separated on individual gels are introduced, and shortcomings of these methods that limit their application in general 2DE analysis are discussed. More approachs including differential in-gel electrophoresis （DIGE）, ＂differential exposure＂approach, and stable isotope approach are stated. The latter methods permitting separation on a single gel containing mixed samples, have superior power in quantitative analysis compared to conventional methods. Various quantitative protocols, advantages and limitations of these protocols as well as further development of quantitative techniques based on 2DE are discussed.