Characterisation of combinatorial libraries of mucin-2 antigen peptides by high-resolution mass spectrometry

Emöke Windberg, Ferenc Hudecz, Andreas Marquardt, Ferenc Sebestyén, Andrea Kiss, Szilvia Bösze, Hedvig Medzihradszky-Schweiger, Michael Przybylski

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

An epitope motif, TX1TX2T, of mucin-2 glycoprotein was identified by means of a mucin-2-specific monoclonal antibody, mAb 994, raised against a synthetic mucin-derived 15-mer peptide conjugate. For determination of the epitope sequence recognised with highest affinity by mAb 994, a combinatorial approach was applied using the portioning-mixing technique excluding Cys. Antibody binding of libraries was most profound when Gln was at the X1 position. Analytical characterisation of the TQTX2T library was conducted by amino acid analysis and matrix-assisted laser desorption/ionisation time-of-flight (MALDI-TOF) and electrospray ionisation Fourier transform ion cyclotron resonance (ESI-FTICR) mass spectrometric methods. Control libraries were prepared by mixing 19 individual peptides corresponding to the TQTX2T sequence. Thus, mixtures of 6, 10 and 19 pentapeptides were analysed and compared with the combinatorial mixture. MALDITOFMS was able to detect only partially the components in the 6- and 10-member mixtures, but failed to characterise a more complex 19-member mixture. In contrast, ESI-FTICRMS resolved all mixtures of higher complexity and provided direct identification at monoisotopic resolution, such as for a peptide library containing 'isobaric' lysine and glutamine (Δm = 0.0364 Da). The results of this study suggest that ESI-FTICRMS is a powerful tool for characterisation of combinatorial peptide libraries of higher complexity.

Original languageEnglish
Pages (from-to)834-839
Number of pages6
JournalRapid Communications in Mass Spectrometry
Volume16
Issue number9
DOIs
Publication statusPublished - May 11 2002

ASJC Scopus subject areas

  • Analytical Chemistry
  • Spectroscopy
  • Organic Chemistry

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