Production of Monoclonal Antibodies Specific for the i and 1,2 Flagellar Antigens of Salmonella typhimurium and Characterization of Their Respective Epitopes

Author:

de Vries N.1,Zwaagstra K. A.12,Huis in’t Veld J. H. J.1,van Knapen F.1,van Zijderveld F. G.3,Kusters J. G.24

Affiliation:

1. Department of the Science of Food of Animal Origin,1 and

2. Department of Bacteriology,2

3. Institute of Infectious Diseases and Immunology, Utrecht University, Utrecht, ID-DLO, Lelystad,3 and

4. Department of Medical Microbiology, Vrije Universiteit, Amsterdam,4 The Netherlands

Abstract

ABSTRACT Salmonella typhimurium expresses two antigenically distinct flagellins, each containing a different H antigen (i and 1,2), the combination of which is highly specific for this serotype. In this study, overlapping recombinant flagellin fragments were constructed from the fliC(H:i) and fljB (H:1,2) flagellin genes, and the expression products were tested for binding to H antigen-specific monoclonal and polyclonal antibodies. A minimal area, 86 amino acids for H:i and 102 amino acids for H:1,2, located in the central variable domain of each flagellin was required for the binding of serotype-specific antibodies, providing further evidence for the presence of a discontinuous H epitope. Two peptides comprising these areas were shown to be highly suitable for application as antigens in an enzyme-linked immunosorbent assay detecting S. typhimurium-specific antibody.

Publisher

American Society for Microbiology

Subject

Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology

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