PRODUCTION OF MONOCLONAL-ANTIBODIES TO THE BORDETELLA-AVIUM 41-KILODALTON SURFACE PROTEIN AND CHARACTERIZATION OF THE HEMAGGLUTININ

Production of monoclonal antibodies (MAbs) to Bordetella avium surface proteins led to further characterization of the hemagglutinin. Protei ns obtained by homogenization of whole-cell B. avium were used to immu nize mice for the production of monoclonal antibodies. Immunoprecipita tion and Western blot techniques were used to determine the specificit y of three MAbs, which all recognized the B. avium 41-kilodalton (kd) surface protein. In addition, all of the MAbs inhibited hemagglutinati on (HA) of guinea pig erythrocytes by B. avium. The 41-kd protein reco gnized by the MAbs was observed by the indirect immunofluorescence tes t and sodium dodecyl sulfate-polyacrylamide gel electrophoresis to bin d to guinea pig erythrocytes. When HA-positive isolates of B. avium we re treated with periodic acid, which cleaves carbohydrates from protei ns, the isolates became HA-negative. Likewise, treatment of HA-positiv e B. avium isolates with proteinase K, which would also remove carbohy drates associated with proteins on the surface of the bacterium, inhib ited HA. Considering these data, we suggest that the B. avium hemagglu tinin is a carbohydrate closely associated with the 41-kd surface prot ein.