Mass spectrometric epitope mapping has become a versatile method to precisely determine a soluble antigen's partial structure that directly interacts with an antibody in solution. Typical lengths of investigated antigens have increased up to several 100 amino acids while experimentally determined epitope peptides have decreased in length to on average 10–15 amino acids. Since the early 1990s more and more sophisticated methods have been developed and have forwarded a bouquet of suitable approaches for epitope mapping with immobilized, temporarily immobilized, and free‐floating antibodies. While up to now monoclonal antibodies have been mostly used in epitope mapping experiments, the applicability of polyclonal antibodies has been proven. The antibody's resistance towards enzymatic proteolysis has been of key importance for the two mostly applied methods: epitope excision and epitope extraction. Sample consumption has dropped to low pmol amounts on both, the antigen and the antibody. While adequate in‐solution sample handling has been most important for successful epitope mapping, mass spectrometric analysis has been found the most suitable read‐out method from early on. The rapidity by which mass spectrometric epitope mapping nowadays is executed outperforms all alternative methods. Thus, it can be asserted that mass spectrometric epitope mapping has reached a state of maturity, which allows it to be used in any mass spectrometry laboratory. After 25 years of constant and steady improvements, its application to clinical samples, for example, for patient characterization and stratification, is anticipated in the near future. © 2016 Wiley Periodicals, Inc. Mass Spec Rev 37:229–241, 2018.

中文翻译：

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质谱表位作图

质谱表位作图已成为一种通用方法，可以精确确定与溶液中的抗体直接相互作用的可溶性抗原的部分结构。被研究抗原的典型长度增加了多达100个氨基酸，而实验确定的表位肽的长度减少了平均10-15个氨基酸。自1990年代初以来，开发了越来越复杂的方法，并提出了一系列适用于固定化，临时固定化和自由浮动抗体的表位作图的合适方法。尽管到目前为止，单克隆抗体已主要用于表位作图实验，但已证明了多克隆抗体的适用性。抗体' 对于两种主要应用的方法：抗原决定簇切除和抗原决定簇提取，其对酶蛋白水解的抵抗力一直至关重要。抗原和抗体的样品消耗均降低至低pmol量。尽管充分的溶液中样品处理对于成功进行表位作图最重要，但从早期开始，人们就发现质谱分析是最合适的读出方法。如今执行质谱表位作图的速度胜过所有其他方法。因此，可以断言质谱表位作图已达到成熟状态，这使其可以在任何质谱实验室中使用。经过25年的不断，稳步改进，它已应用于临床样本，例如，用于患者表征和分层，预计在不久的将来。©2016 Wiley Periodicals，Inc.质谱修订37：229–241，2018。