Hager-Braun Christine, Tomer Kenneth B
Laboratory of Structural Biology, National Institute of Environmental Health Sciences, NIH, DHHS, Research Triangle Park, NC 27709, USA.
Expert Rev Proteomics. 2005 Oct;2(5):745-56. doi: 10.1586/14789450.2.5.745.
Mass spectrometry has evolved as a technique suitable for the characterization of peptides and proteins beyond their linear sequence. The advantages of mass spectrometric sample analysis are high sensitivity, high mass accuracy, rapid analysis time and low sample consumption. In epitope mapping, the molecular structure of an antigen (the epitope or antigenic determinant) that interacts with the paratope (recognition surface) of the antibody is identified. To obtain information on linear, conformational and/or discontinuous epitopes, various approaches have been developed in conjunction with mass spectrometry. These methods include limited proteolysis and epitope footprinting, epitope excision and epitope extraction for linear epitopes and probing the surface accessibility of residues by differential chemical modifications of specific amino acid side chains or by differential hydrogen/deuterium exchange of the protein backbone amides for conformational and discontinuous epitopes. Epitope mapping by mass spectrometry is applicable in basic biochemical research and, with increasing robustness, should soon find its implementation in routine clinical diagnosis.
质谱已发展成为一种适用于表征肽和蛋白质线性序列之外特征的技术。质谱样品分析的优点是灵敏度高、质量精度高、分析时间快和样品消耗低。在表位作图中，可识别与抗体的互补位（识别表面）相互作用的抗原（表位或抗原决定簇）的分子结构。为了获得关于线性、构象和/或不连续表位的信息，已结合质谱开发了各种方法。这些方法包括用于线性表位的有限蛋白酶解和表位足迹分析、表位切除和表位提取，以及通过特定氨基酸侧链的差异化学修饰或通过蛋白质主链酰胺的差异氢/氘交换来探测残基的表面可及性，以确定构象和不连续表位。通过质谱进行表位作图适用于基础生化研究，并且随着稳健性的提高，很快将在常规临床诊断中得到应用。
