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Permanent URL to this publication: http://dx.doi.org/10.5167/uzh-13773

Pimenova, T; Nazabal, A; Roschitzki, B; Seebacher, J; Rinner, O; Zenobi, R (2008). Epitope mapping on bovine prion protein using chemical cross-linking and mass spectrometry. Journal of Mass Spectrometry, 43(2):185-195.

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Abstract

An analytical strategy for the analysis of antigen epitopes by chemical cross-linking and mass 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.

Item Type:Journal Article, refereed, original work
Communities & Collections:04 Faculty of Medicine > Functional Genomics Center Zurich
08 University Research Priority Programs > Systems Biology / Functional Genomics
DDC:570 Life sciences; biology
610 Medicine & health
Language:English
Date:2008
Deposited On:19 Feb 2009 11:22
Last Modified:27 Nov 2013 17:49
Publisher:Wiley-Blackwell
ISSN:1076-5174
Free access at:Publisher DOI. An embargo period may apply.
Publisher DOI:10.1002/jms.1280
PubMed ID:17924399
Citations:Web of Science®. Times Cited: 21
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