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Infection and Immunity, July 2000, p. 3949-3955, Vol. 68, No. 7
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Selection of Recombinant Antibodies Specific for Pathogenic Streptococcus suis by Subtractive Phage Display

Astrid de Greeff,1,2,* Loek van Alphen,1,3 and Hilde E. Smith2

Department of Medical Microbiology, University of Amsterdam, Academical Medical Center, Amsterdam,1 Department of Bacteriology, Institute of Animal Science and Health, 8200 AB Lelystad,2 and Laboratory of Vaccine Development and Immune Mechanisms, RIVM, National Institute of Public Health and the Environment, 3720 BA Bilthoven,3 The Netherlands

Received 6 January 2000/Returned for modification 14 February 2000/Accepted 23 April 2000

A semisynthetic antibody phage display library was used to select recombinant antibodies directed against surface components of a pathogenic strain of Streptococcus suis serotype 2 and against extracellular factor (EF), a protein known to be exclusively associated with pathogenic S. suis serotype 2 strains. Three distinct monoclonal phage antibodies directed against conformational epitopes of surface protein components of S. suis were selected. In addition, three different monoclonal phage antibodies were isolated that recognized EF. To isolate antibody fragments that recognize epitopes specific for a pathogenic S. suis serotype 2 strain, compared to a nonpathogenic serotype 2 strain, we applied a subtractive selection procedure. With this procedure, only one distinct phage antibody was found, and it was shown to be directed against EF. This demonstrates the selectivity of the applied procedure and confirms that EF is indeed differentially expressed by pathogenic and nonpathogenic strains. It also shows that EF is a very dominant antigen in phage antibody selections.

* Corresponding author. Mailing address: Department of Bacteriology, Institute of Animal Science and Health, P.O. Box 65, 8200 AB Lelystad, The Netherlands. Phone: 31 320 238403. Fax: 31 320 238153. E-mail: a.degreeff{at}id.wag-ur.nl.

Infection and Immunity, July 2000, p. 3949-3955, Vol. 68, No. 7
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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