Department of Immunotechnology, Lund University, Lund, Sweden, and CREATE Health, BMC D13, Lund, Sweden.
Mol Cell Proteomics. 2011 Oct;10(10):M110.003962. doi: 10.1074/mcp.M110.003962. Epub 2011 Jun 14.
Antibody-based microarrays are a rapidly evolving affinity-proteomic methodology that recently has shown great promise in clinical applications. The resolution of these proteomic analyses is, however, directly related to the number of data-points, i.e. antibodies, included on the array. Currently, this is a key bottleneck because of limited availability of numerous highly characterized antibodies. Here, we present a conceptually new method, denoted global proteome survey, opening up the possibility to probe any proteome in a species-independent manner while still using a limited set of antibodies. We use context-independent-motif-specific antibodies directed against short amino acid motifs, where each motif is present in up to a few hundred different proteins. First, the digested proteome is exposed to these antibodies, whereby motif-containing peptides are enriched, which then are detected and identified by mass spectrometry. In this study, we profiled extracts from human colon tissue, yeast cells lysate, and mouse liver tissue to demonstrate proof-of-concept.
基于抗体的微阵列是一种快速发展的亲和蛋白质组学方法，最近在临床应用中显示出巨大的潜力。然而，这些蛋白质组学分析的分辨率与数组上包含的数据点（即抗体）的数量直接相关。目前，由于大量高度表征的抗体的可用性有限，这是一个关键的瓶颈。在这里，我们提出了一种全新的方法，称为全局蛋白质组调查，为在不依赖于物种的情况下探测任何蛋白质组提供了可能性，同时仍然使用有限数量的抗体。我们使用针对短氨基酸基序的不依赖于上下文的基序特异性抗体，其中每个基序存在于多达几百种不同的蛋白质中。首先，将消化的蛋白质组暴露于这些抗体，使含有基序的肽被富集，然后通过质谱检测和鉴定。在这项研究中，我们对人结肠组织、酵母细胞裂解物和小鼠肝组织的提取物进行了分析，以证明其概念验证。
