Advanced Biotechnology Center, Largo Rosanna Benzi, Genova, Italy.
Proteomics. 2010 Oct;10(20):3566-76. doi: 10.1002/pmic.201000245.
Over the last years, top-down (TD) MS has gained a remarkable space in proteomics, rapidly trespassing the limit between a promising approach and a solid, established technique. Several research groups worldwide have implemented TD analysis in their routine work on proteomics, deriving structural information on proteins with the level of accuracy that is impossible to achieve with classical bottom-up approaches. Complete maps of PTMs and assessment of single aminoacid polymorphisms are only a few of the results that can be obtained with this technique. Despite some existing technical and economical limitations, TD analysis is at present the most powerful instrument for MS-based proteomics and its implementation in routine workflow is a rapidly approaching turning point in proteomics. In this review article, the state-of-the-art of TD approach is described along with its major advantages and drawbacks and the most recent trends in TD analysis are discussed. References for all the covered topics are reported in the text, with the aim to support both newcomers and mass spectrometrists already introduced to TD proteomics.
近年来，自上而下（TD）MS 在蛋白质组学中获得了显著的发展空间，迅速超越了有前途的方法和成熟、确立的技术之间的界限。全球有几个研究小组在他们的蛋白质组学常规工作中实施了 TD 分析，获得了蛋白质结构信息，其准确性是经典的自下而上方法无法实现的。完整的 PTM 图谱和单个氨基酸多态性的评估只是该技术可以获得的结果中的一小部分。尽管存在一些现有的技术和经济限制，但 TD 分析目前是基于 MS 的蛋白质组学最强大的工具，其在常规工作流程中的实施是蛋白质组学的一个迅速逼近的转折点。在这篇综述文章中，描述了 TD 方法的最新进展，包括其主要优点和缺点，以及 TD 分析的最新趋势。文本中报告了所有涵盖主题的参考文献，旨在为新入门者和已经熟悉 TD 蛋白质组学的质谱仪技术人员提供支持。
