Yu Xiaobo, Xu Danke, Cheng Quan
Department of Biochemistry, Beijing Institute of Radiation Medicine, Beijing Proteome Research Center, Beijing, PR China.
Proteomics. 2006 Oct;6(20):5493-503. doi: 10.1002/pmic.200600216.
With the growth of the "-omics" such as functional genomics and proteomics, one of the foremost challenges in biotechnologies has become the development of novel methods to monitor biological process and acquire the information of biomolecular interactions in a systematic manner. To fully understand the roles of newly discovered genes or proteins, it is necessary to elucidate the functions of these molecules in their interaction network. Microarray technology is becoming the method of choice for such a task. Although protein microarray can provide a high throughput analytical platform for protein profiling and protein-protein interaction, most of the current reports are limited to labeled detection using fluorescence or radioisotope techniques. These limitations deflate the potential of the method and prevent the technology from being adapted in a broader range of proteomics applications. In recent years, label-free analytical approaches have gone through intensified development and have been coupled successfully with protein microarray. In many examples of label-free study, the microarray has not only offered the high throughput detection in real time, but also provided kinetics information as well as in situ identification. This article reviews the most significant label-free detection methods for microarray technology, including surface plasmon resonance imaging, atomic force microscope, electrochemical impedance spectroscopy and MS and their applications in proteomics research.
随着功能基因组学和蛋白质组学等“组学”技术的发展，生物技术领域最主要的挑战之一已变成开发新方法，以系统地监测生物过程并获取生物分子相互作用的信息。为了全面理解新发现基因或蛋白质的作用，有必要阐明这些分子在其相互作用网络中的功能。微阵列技术正成为完成这项任务的首选方法。尽管蛋白质微阵列可为蛋白质谱分析和蛋白质-蛋白质相互作用提供高通量分析平台，但目前大多数报告仅限于使用荧光或放射性同位素技术进行标记检测。这些局限性削弱了该方法的潜力，阻碍了该技术在更广泛的蛋白质组学应用中的采用。近年来，无标记分析方法得到了深入发展，并已成功与蛋白质微阵列相结合。在许多无标记研究实例中，微阵列不仅提供了实时高通量检测，还提供了动力学信息以及原位鉴定。本文综述了微阵列技术最重要的无标记检测方法，包括表面等离子体共振成像、原子力显微镜、电化学阻抗谱和质谱及其在蛋白质组学研究中的应用。
