Department of Biosciences and Bioengineering, Indian Institute of Technology Bombay, Mumbai, India.
Proteomics. 2010 Feb;10(4):731-48. doi: 10.1002/pmic.200900458.
Protein microarrays, on which thousands of discrete proteins are printed, provide a valuable platform for functional analysis of the proteome. They have been widely used for biomarker discovery and to study protein-protein interactions. The accomplishments of DNA microarray technology, which had enabled massive parallel studies of gene expression, sparked great interest for the development of protein microarrays to achieve similar success at the protein level. Protein microarray detection techniques are often classified as being label-based and label-free. Most of the microarray applications have employed labelled detection such as fluorescent, chemiluminescent and radioactive labelling. These labelling strategies have synthetic challenges, multiple label issues and may exhibit interference with the binding site. Therefore, development of sensitive, reliable, high-throughput, label-free detection techniques are now attracting significant attention. Label-free detection techniques monitor biomolecular interactions and simplify the bioassays by eliminating the need for secondary reactants. Moreover, they provide quantitative information for the binding kinetics. In this article, we will review several label-free techniques, which offer promising applications for the protein microarrays, and discuss their prospects, merits and challenges.
蛋白质微阵列在其上打印了数千种离散的蛋白质,为蛋白质组的功能分析提供了有价值的平台。它们已被广泛用于生物标志物的发现和研究蛋白质-蛋白质相互作用。DNA 微阵列技术的成就使得大规模并行研究基因表达成为可能,这激发了人们极大的兴趣,希望开发出蛋白质微阵列,在蛋白质水平上取得类似的成功。蛋白质微阵列检测技术通常分为基于标记和无标记两种。大多数微阵列应用都采用了标记检测,如荧光、化学发光和放射性标记。这些标记策略具有合成挑战、多个标记问题,并且可能会干扰结合位点。因此,现在人们对开发敏感、可靠、高通量、无标记的检测技术产生了浓厚的兴趣。无标记检测技术通过消除对二级反应物的需求来监测生物分子相互作用并简化生物测定。此外,它们还提供了有关结合动力学的定量信息。本文将综述几种无标记技术,它们为蛋白质微阵列提供了有前途的应用,并讨论它们的前景、优点和挑战。