The Ben May Department for Cancer Research, and The Institute for Genomics & Systems Biology, The University of Chicago, Chicago, IL 60637, USA.
Expert Rev Proteomics. 2011 Oct;8(5):565-75. doi: 10.1586/epr.11.49.
While proteomic methods have illuminated many areas of biological protein space, many fundamental questions remain with regard to systems-level relationships between mRNAs, proteins and cell behaviors. While mass spectrometric methods offer a panoramic picture of the relative expression and modification of large numbers of proteins, they are neither optimal for the analysis of predefined targets across large numbers of samples nor for assessing differences in proteins between individual cells or cell compartments. Conversely, traditional antibody-based methods are effective at sensitively analyzing small numbers of proteins across small numbers of conditions, and can be used to analyze relative differences in protein abundance and modification between cells and cell compartments. However, traditional antibody-based approaches are not optimal for analyzing large numbers of protein abundances and modifications across many samples. In this article, we will review recent advances in methodologies and philosophies behind several microarray-based, intermediate-level, 'protein-omic' methods, including a focus on reverse-phase lysate arrays and micro-western arrays, which have been helpful for bridging gaps between large- and small-scale protein analysis approaches and have provided insight into the roles that protein systems play in several biological processes.
虽然蛋白质组学方法已经阐明了生物蛋白质空间的许多领域,但关于 mRNA、蛋白质和细胞行为之间的系统水平关系仍有许多基本问题。虽然质谱方法提供了大量蛋白质相对表达和修饰的全景图,但它们既不适合分析大量样本中预定义的靶标,也不适合评估单个细胞或细胞区室之间蛋白质的差异。相反,传统的基于抗体的方法在分析少数条件下的少数蛋白质方面非常有效,并且可用于分析细胞和细胞区室之间蛋白质丰度和修饰的相对差异。然而,传统的基于抗体的方法并不适合分析许多样本中大量蛋白质的丰度和修饰。在本文中,我们将回顾几种基于微阵列的、中等水平的“蛋白质组学”方法背后的方法学和哲学的最新进展,包括重点介绍反相裂解物阵列和微西方阵列,这些方法有助于弥合大规模和小规模蛋白质分析方法之间的差距,并深入了解蛋白质系统在几个生物学过程中所起的作用。