Selection of recombinant antibodies specific for pathogenic Streptococcus suis by subtractive phage display.
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.
de Greeff
A
A
Department of Medical Microbiology, University of Amsterdam, Academical Medical Center, Amsterdam, The Netherlands. a.degreeff@id.wag-ur.nl
van Alphen
L
L
Smith
H E
HE
D000595:Amino Acid Sequence; D000907:Antibodies, Bacterial / Q000235:genetics* / Q000302:isolation & purification; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000302:isolation & purification; D000916:Antibody Diversity; D000918:Antibody Specificity; D000942:Antigens, Bacterial; D001426:Bacterial Proteins / Q000276:immunology; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D000939:Epitopes; D006801:Humans; D019151:Peptide Library; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D017011:Streptococcus suis / Q000235:genetics / Q000276:immunology* / Q000472:pathogenicity
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D017931:DNA Primers; C082316:Epf protein, Streptococcus suis; D000939:Epitopes; D019151:Peptide Library; D011994:Recombinant Proteins