Proteomic Unit, CIC bioGUNE, CIBERehd, ProteoRed, Derio, Spain.
Mass Spectrom Rev. 2013 Jan-Feb;32(1):1-26. doi: 10.1002/mas.21347. Epub 2012 Jul 30.
The systematic analysis of biological processes requires an understanding of the quantitative expression patterns of proteins, their interacting partners and their subcellular localization. This information was formerly difficult to accrue as the relative quantification of proteins relied on antibody-based methods and other approaches with low throughput. The advent of soft ionization techniques in mass spectrometry plus advances in separation technologies has aligned protein systems biology with messenger RNA, DNA, and microarray technologies to provide data on systems as opposed to singular protein entities. Another aspect of quantitative proteomics that increases its importance for the coming few years is the significant technical developments underway both for high pressure liquid chromatography and mass spectrum devices. Hence, robustness, reproducibility and mass accuracy are still improving with every new generation of instruments. Nonetheless, the methods employed require validation and comparison to design fit for purpose experiments in advanced protein analyses. This review considers the newly developed systematic protein investigation methods and their value from the standpoint that relative or absolute protein quantification is required de rigueur in biomedical research.
系统分析生物过程需要了解蛋白质、其相互作用伙伴及其亚细胞定位的定量表达模式。由于相对蛋白质定量依赖于抗体方法和其他低通量方法,因此以前很难获得这些信息。质谱中的软电离技术的出现以及分离技术的进步,使蛋白质系统生物学与信使 RNA、DNA 和微阵列技术相结合,提供了有关系统的信息,而不是单个蛋白质实体的信息。定量蛋白质组学的另一个方面是,它在未来几年的重要性日益增加,因为高压液相色谱和质谱设备都在进行重大技术开发。因此,每一代新仪器的稳健性、可重复性和质量精度都在不断提高。尽管如此,所采用的方法仍需要验证和比较,以设计适合目的的实验,进行高级蛋白质分析。从相对或绝对蛋白质定量是生物医学研究的必要条件的角度来看,本文综述了新开发的系统蛋白质研究方法及其价值。