title: Mass spectrometric epitope mapping.
abstract: 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.
authors.affiliation: Proteome Center Rostock, University Medicine and Natural Science Faculty, University of Rostock, Rostock, Germany.
authors.forename: Kwabena F M
authors.initials: KFM
authors.lastname: Opuni
authors.affiliation: Proteome Center Rostock, University Medicine and Natural Science Faculty, University of Rostock, Rostock, Germany.
authors.forename: Mahmoud
authors.initials: M
authors.lastname: Al-Majdoub
authors.affiliation: Proteome Center Rostock, University Medicine and Natural Science Faculty, University of Rostock, Rostock, Germany.
authors.forename: Yelena
authors.initials: Y
authors.lastname: Yefremova
authors.affiliation: Microbiology and Immunology Faculty of Pharmacy, The British University in Egypt, Cairo, Egypt.
authors.forename: Reham F
authors.initials: RF
authors.lastname: El-Kased
authors.affiliation: Proteome Center Rostock, University Medicine and Natural Science Faculty, University of Rostock, Rostock, Germany.
authors.forename: Cornelia
authors.initials: C
authors.lastname: Koy
authors.affiliation: Proteome Center Rostock, University Medicine and Natural Science Faculty, University of Rostock, Rostock, Germany.
authors.forename: Michael O
authors.initials: MO
authors.lastname: Glocker
mesh_terms: D000818:Animals; D055910:Antibodies, Immobilized / Q000737:chemistry; D018604:Epitope Mapping / Q000379:methods*; D000939:Epitopes / Q000302:isolation & purification; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*
chemical_list: D055910:Antibodies, Immobilized; D000939:Epitopes
keywords: assembled epitope; conformational epitope; consecutive epitope; discontinuous epitope; epitope excision; epitope extraction; linear epitope; mass spectrometry