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.

Systems vaccinology and big data in the vaccine development chain

“…5 However, the role of immunogens is currently studied in conjunction with other factors, such as adjuvants and antigen delivery, as these influence immune responses and induction of immunological memory that are key for vaccine ª 2018 The Authors. Immunology Published by John Wiley & Sons Ltd., Immunology, 156, [33][34][35][36][37][38][39][40][41][42][43][44][45][46] efficacy. Systems vaccinology addresses these aspects in order to understand why some vaccines function properly whereas other vaccines do not.…”

“…With instrumental improvement of MS and the supporting software tools, MS is now used as a versatile tool to determine B-cell epitopes, that is, the areas where antibodies bind. Opuni et al 33 comprehensively reviewed MS-based approaches to map B-cell epitopes. Antibody-antigen binding is just one example of a protein-protein interaction.…”

Epitope Mapping of Antibody-Antigen Interactions with X-Ray Crystallography

“…A variety of techniques are available for epitope mapping including: peptide display technologies (Pepscan) [6], NMR spectroscopy [7], mass spectrometry [8], and X-ray crystallography. While each of these methods has their own advantages and drawbacks, one issue to consider is the specific natures of the epitope recognized by the antibody.…”

Native Mass Spectrometry, Ion mobility, and Collision-Induced Unfolding Categorize Malaria Antigen/Antibody Binding

“…Owing to its sequencing capability, MS is one “read-out” for many functional epitope-mapping approaches [2, 3]. Moreover, MS-based footprinting (e.g., HDX-MS) have successfully bridged high and low-resolution structural epitope mapping methods by reliably localizing the binding site at the peptide level [4–8].…”