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Abstract No.: ThOr-25   [KEYNOTE LECTURE]
Speaker: Michael Przybylski
Session: MS Contribution to Immunology
Presentation date: Thu, Aug 31, 2006
Presentation time: 16:00 – 16:30

Mass Spectrometric Approaches for Elucidation of Antigen-antibody Recognition Structures in Molecular Immunology

Michael Przybylski1, Raluca Stefanescu1, Michael Bacher2, Marilena Manea1, Irina Perdivara1, Madalina Maftei1, Gabriela Paraschiv1, Erika R. Amstalden van Hove1, Andreas Marquardt1, Richard Dodel2

1 University of Konstanz, Konstanz, Germany
2 University of Bonn, Bonn, Germany

Correspondence address: Michael Przybylski, University of Konstanz, Department of Chemistry, Laboratory of Analytical Chemistry, Universitaetsstrasse 10, Konstanz, 78457 Germany.

Web site: http://www.uni-konstanz.de/agprzybylski/chemie

Keywords: Affinity, Binding; Antibodies; Antigens, Peptide; Fourier Transform ICR; Molecular Recognition; Protein Identification; Proteomic.

In the last years, mass spectrometric approaches have gained increasing access to molecular immunology, and developed for chemical structure identifications of antigen-antibody interactions.1-3 Using high resolution Fouriertransform ion cyclotron resonance mass spectrometry (FTICR MS) as a powerful proteomics tool with unrivalled accuracy, recent work in our laboratory has focussed on the development of high resolution and high selectivity MS approaches, and applications to the identification of antibody recognition structures, as a key pre-requisite for vaccine design and targeting. Selective proteolytic digestion and MS-peptide mapping (EPITOPE EXCISION) has been successfully employed for epitope identification of protein antigens. In addition, "affinity-proteomics" using partial epitope excision has been developed as an approach with unprecedented selectivity for direct protein identification from biological material.2,3 The potential of these methods has been demonstrated by the elucidation of a β-amyloid-plaque-specific epitope recognised by therapeutic antibodies from transgenic mouse models of Alzheimer`s disease.4 Using this epitope in an antigen-affinity column, and antibody-proteolytic digestion and analysis by FTICR MS, a new approach has been developed for the identification of antibody recognition structures (paratope-proteomics; “[PAREXPROT]”). In this method, high resolution MS-peptide mapping data at the 1 ppm level are applied for the identification of paratopes from protein databases.5 First applications to the elucidation of paratope-containing peptides within heavy- and light-chain fragments of β-amyloid- antibodies specific to Alzheimer’s disease will be discussed. Mass spectrometric epitope mapping, and antibody-proteomics for determination of "molecular antibody-recognition signatures" offer high perspectives for the development of new molecular diagnostics, receptor structural characterisation, and the evaluation of lead structures for new vaccines.

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5. M. Przybylski, E. Amstalden, A. Marquardt, R. Iacob, R. Stefanescu, I. Perdivara and M. Manea, Nature Meth. submitted (2006).