Epitope mapping on bovine prion protein using chemical cross-linking and mass spectrometry

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Author(s) Pimenova, Tatiana, Nazabal, Alexis, Roschitzki, Bernd, Seebacher, Jan, Rinner, Oliver, Zenobi, Renato
Publication Type Journal Items, Publication Status: Published
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Title Epitope mapping on bovine prion protein using chemical cross-linking and mass spectrometry
Author(s) Pimenova, Tatiana
Nazabal, Alexis
Roschitzki, Bernd
Seebacher, Jan
Rinner, Oliver
Zenobi, Renato
Journal or Series Title Journal of mass spectrometry
Volume Number 43
Issue Number 2
Start Page 185
End Page 195
ISSN 1076-5174
Publisher Wiley
Publication Place Chichester
Publication Date 2008
Keyword(s) antigen/antibody complex
mass spectrometry
epitope mapping
high-mass MALDI-TOF MS
Abstract An analytical strategy for the analysis of antigen epitopes by chemical cross-linking andmass spectrometry is demonstrated. The information of antigen peptides involved in the binding to an antibody can be obtained by monitoring the antigen peptides modified by a partially hydrolyzed cross-linker in the absence and in the presence of an antibody. This approach was shown to be efficient for characterization of the epitope on bovine prion protein bPrP(25–241) specifically recognized by a monoclonal antibody, 3E7 (mAb3E7), with only a small amount of sample (200 picomoles) needed. After cross-linking of the specific immuno complex, a matrix-assisted laser desorption/ionization (MALDI) mass spectrometer equipped with an ion conversion dynode (ICD) high-mass detector was used to optimize the amount of cross-linked complex formed at 202 kDa before proteolytic digestion. To identify the cross-linked peptides after proteolysis without ambiguity, isotope-labeled cross-linkers, disuccinimidyl suberate (DSS-d0/d12) and disuccinimidyl glutarate (DSG-d0/d6), together with high-resolution Fourier transform ion-cyclotron resonance mass spectrometry (FTICR-MS) were used. As a result, a complete fading of the peak intensities corresponding to the peptides representing the epitope was observed when bPrP/mAb3E7 complexes were formed.
DOI 10.1002/jms.1280
Additional Notes Received 7 May 2007, Accepted 9 July 2007, Published online 8 October 2007
Document Type Article
Publication Status Published
Language English
Assigned Organisational Unit(s) 02207
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NEBIS System Number 001410098
Source Database ID PP-40961
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  author = "Pimenova, Tatiana and Nazabal, Alexis and Roschitzki, Bernd and Seebacher, Jan and Rinner, Oliver and Zenobi, Renato",
  title = "{E}pitope mapping on bovine prion protein using chemical cross-linking and mass spectrometry",
  journal = "Journal of mass spectrometry",
  year = 2008,
  volume = "43",
  number = "2",
  pages = "185--195",

E-Citations record created: Thu, 01 Apr 2010, 22:43:57 CET