Intact Transition Epitope Mapping (ITEM)

Yelena Yefremova, Kwabena F.M. Opuni, Bright D. Danquah, Hans Juergen Thiesen, Michael O. Glocker

Research output: Contribution to journalArticlepeer-review

24 Citations (Scopus)

Abstract

Intact transition epitope mapping (ITEM) enables rapid and accurate determination of protein antigen-derived epitopes by either epitope extraction or epitope excision. Upon formation of the antigen peptide-containing immune complex in solution, the entire mixture is electrosprayed to translate all constituents as protonated ions into the gas phase. There, ions from antibody–peptide complexes are separated from unbound peptide ions according to their masses, charges, and shapes either by ion mobility drift or by quadrupole ion filtering. Subsequently, immune complexes are dissociated by collision induced fragmentation and the ion signals of the “complex-released peptides,” which in effect are the epitope peptides, are recorded in the time-of-flight analyzer of the mass spectrometer. Mixing of an antibody solution with a solution in which antigens or antigen-derived peptides are dissolved is, together with antigen proteolysis, the only required in-solution handling step. Simplicity of sample handling and speed of analysis together with very low sample consumption makes ITEM faster and easier to perform than other experimental epitope mapping methods.

Original languageEnglish
Pages (from-to)1612-1622
Number of pages11
JournalJournal of the American Society for Mass Spectrometry
Volume28
Issue number8
DOIs
Publication statusPublished - 1 Aug 2017
Externally publishedYes

Keywords

  • Antibody–antigen interactions
  • Antibody–epitope reactivities
  • Ion mobility separation
  • Native electrospray mass spectrometry
  • Quadrupole time-of-flight mass spectrometry

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