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Identification and mapping of a linear epitope of centromere protein F using monoclonal antibodies

Authors

  • Simon Welner,

    1. Department of Clinical Biochemistry, Immunology and Genetics, Statens Serum Institut, Copenhagen S, Denmark
    2. IGM – Bioorganic Chemistry, Faculty of Life Sciences, University of Copenhagen, Frederiksberg C, Denmark
    3. Present address: LEO Pharma A/S, Ballerup, Denmark
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  • Nicole H. Trier,

    1. Department of Clinical Biochemistry, Immunology and Genetics, Statens Serum Institut, Copenhagen S, Denmark
    2. IGM – Bioorganic Chemistry, Faculty of Life Sciences, University of Copenhagen, Frederiksberg C, Denmark
    3. Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen Ø, Denmark
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  • Gunnar Houen,

    1. Department of Clinical Biochemistry, Immunology and Genetics, Statens Serum Institut, Copenhagen S, Denmark
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  • Paul R. Hansen

    Corresponding author
    1. IGM – Bioorganic Chemistry, Faculty of Life Sciences, University of Copenhagen, Frederiksberg C, Denmark
    • Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen Ø, Denmark
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Correspondence to: Paul R. Hansen, Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, 2100 Copenhagen Ø, Denmark. E-mail: prh@sund.ku.dk

Abstract

Autoantibodies against centromere protein -F have been reported to be associated with various types of cancer with poor prognosis. The characterization of these autoantibody specificities is important in both diagnostics and basic research. In this study, we mapped the epitope (NELSRIRSEKA) of two monoclonal centromere protein F antibodies. The epitope was localized by screening of overlapping peptides followed by a fast and efficient estimation of the minimal peptide length required for antibody recognition, based on the screening of terminally truncated resin-bound peptide analogs. The epitope was determined through competitive inhibition assays of systematically truncated free peptides. In addition, the importance of the involved amino acid side chains of the identified epitope was determined through competitive inhibition assays using alanine-substituted analogs. Copyright © 2013 European Peptide Society and John Wiley & Sons, Ltd.

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