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

Emöke Windberg, F. Hudecz, Andreas Marquardt, Ferenc Sebestyén, Andrea Kiss, Sz. Bősze, 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 - 2002

Fingerprint

Mucin-2
Mass spectrometry
Antigens
Peptides
Peptide Library
Epitopes
Cyclotron resonance
Electrospray ionization
Mucins
Glutamine
Lysine
Ionization
Desorption
Glycoproteins
Fourier transforms
Monoclonal Antibodies
Ions
Amino Acids
Antibodies
Lasers

ASJC Scopus subject areas

  • Analytical Chemistry
  • Spectroscopy

Cite this

Characterisation of combinatorial libraries of mucin-2 antigen peptides by high-resolution mass spectrometry. / Windberg, Emöke; Hudecz, F.; Marquardt, Andreas; Sebestyén, Ferenc; Kiss, Andrea; Bősze, Sz.; Medzihradszky-Schweiger, Hedvig; Przybylski, Michael.

In: Rapid Communications in Mass Spectrometry, Vol. 16, No. 9, 2002, p. 834-839.

Research output: Contribution to journalArticle

Windberg, E, Hudecz, F, Marquardt, A, Sebestyén, F, Kiss, A, Bősze, S, Medzihradszky-Schweiger, H & Przybylski, M 2002, 'Characterisation of combinatorial libraries of mucin-2 antigen peptides by high-resolution mass spectrometry', Rapid Communications in Mass Spectrometry, vol. 16, no. 9, pp. 834-839. https://doi.org/10.1002/rcm.649
Windberg, Emöke ; Hudecz, F. ; Marquardt, Andreas ; Sebestyén, Ferenc ; Kiss, Andrea ; Bősze, Sz. ; Medzihradszky-Schweiger, Hedvig ; Przybylski, Michael. / Characterisation of combinatorial libraries of mucin-2 antigen peptides by high-resolution mass spectrometry. In: Rapid Communications in Mass Spectrometry. 2002 ; Vol. 16, No. 9. pp. 834-839.
@article{c8c166160dd74ee58470fc69ae6db95a,
title = "Characterisation of combinatorial libraries of mucin-2 antigen peptides by high-resolution mass spectrometry",
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.",
author = "Em{\"o}ke Windberg and F. Hudecz and Andreas Marquardt and Ferenc Sebesty{\'e}n and Andrea Kiss and Sz. Bősze and Hedvig Medzihradszky-Schweiger and Michael Przybylski",
year = "2002",
doi = "10.1002/rcm.649",
language = "English",
volume = "16",
pages = "834--839",
journal = "Rapid Communications in Mass Spectrometry",
issn = "0951-4198",
publisher = "John Wiley and Sons Ltd",
number = "9",

}

TY - JOUR

T1 - Characterisation of combinatorial libraries of mucin-2 antigen peptides by high-resolution mass spectrometry

AU - Windberg, Emöke

AU - Hudecz, F.

AU - Marquardt, Andreas

AU - Sebestyén, Ferenc

AU - Kiss, Andrea

AU - Bősze, Sz.

AU - Medzihradszky-Schweiger, Hedvig

AU - Przybylski, Michael

PY - 2002

Y1 - 2002

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0036010939&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036010939&partnerID=8YFLogxK

U2 - 10.1002/rcm.649

DO - 10.1002/rcm.649

M3 - Article

C2 - 11948813

AN - SCOPUS:0036010939

VL - 16

SP - 834

EP - 839

JO - Rapid Communications in Mass Spectrometry

JF - Rapid Communications in Mass Spectrometry

SN - 0951-4198

IS - 9

ER -