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Applied and Environmental Microbiology, December 1998, p. 5033-5038, Vol. 64, No. 12
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

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

N. de Vries,1 K. A. Zwaagstra,1,2 J. H. J. Huis in't Veld,1 F. van Knapen,1 F. G. van Zijderveld,3 and J. G. Kusters2,4,*

Department of the Science of Food of Animal Origin,1 and Department of Bacteriology,2 Institute of Infectious Diseases and Immunology, Utrecht University, Utrecht, ID-DLO, Lelystad,3 and Department of Medical Microbiology, Vrije Universiteit, Amsterdam,4 The Netherlands

Received 7 April 1998/Accepted 25 August 1998

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.

* Corresponding author. Mailing address: Department of Medical Microbiology, Vrije Universiteit, Van der Boechorststraat 7, 1081 BT, Amsterdam, The Netherlands. Phone: (31) 20 4448310. Fax: (31) 20 4448318. E-mail: jg.kusters.mm{at}med.vu.nl.

Applied and Environmental Microbiology, December 1998, p. 5033-5038, Vol. 64, No. 12
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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