[Rapid purification of Bordetella pertussis fimbria 2, 3 proteins and their immunogenicities].
Bordetella pertussis fimbria antigens play an important role in the adhesion to the cell surfaces, and induce the formation of protective antibodies in the host. The aim of this study was to purify the fimbria proteins by a rapid and easy method which involved Superdex 75 gel filtration chromatography, and to investigate their immunogenicity. The fimbria proteins purified from the different strains showed single protein bands in 12.5% SDS-PAGE (Sodium dodecyl sulphate-polyacrylamide gel electrophoresis) method. Depending on the strain used, the proteins were observed as Fim2 and Fim3 subunits or only Fim 2 subunit on 16% SDS-PAGE. The purified subunits were confirmed by Western and dot blotting analysis with monoclonal antibodies BP F2 (anti-Fim2) and BPC10 (anti-Fim 3). The immunogenicity studies performed in the mice revealed that purified fimbria proteins have led to higher agglutinin response than the vaccine with whole cell pertussis antigens. It was determined that the extent of agglutinin responses were different for different strains from which the fimbria proteins were purified. In addition, each B. pertussis strain showed a different agglutination reaction with different anti-Fim sera. The results, overall, pointed out the significance of fimbria structure of the B. pertussis strain used for vaccine manufacture and control.
Atakan Ablay
Ptnar
P
Refik Saydam Hifzissihha Merkezi Başkanliği, Aşi-Serum Uretim ve Araştirma Bölümü, Ankara.
Ozcengiz
Erkan
E
D000372:Agglutination Tests; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000942:Antigens, Bacterial / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D015153:Blotting, Western; D001886:Bordetella pertussis / Q000276:immunology*; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D036781:Fimbriae Proteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D010861:Fimbriae, Bacterial / Q000276:immunology*; D015151:Immunoblotting; D051379:Mice
D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D036781:Fimbriae Proteins