Production and Characterization of Monoclonal Antibodies to Serotype Specific Antigens of Haemophilus Pleuropneumoniae Serotype 2

Abstract

Mouse monoclonal antibodies were produced to Haemophilus pleuropneumoniae bacteria of the most common serotype 2. 11 hybridoma cultures were recovered that produced antibodies with moderate to strong reactivity to the antigen. 10 of these antibodies were specific to isolated capsular antigens from H. pleuropneumoniae of serotype 2, while one antibody reacted with capsular antigens from bacteria of all 8 serotypes. One hybridoma producing antibodies with a titre of 1:1000 was cloned and the antibody specificity studied further. The binding of this antibody (1F3) to whole bacteria, and capsular extracts isolated at different temperatures indicate that the antibody is specific for a thermostable polysaccharide antigen present in the cellular capsule of H. pleuropneumoniae of serotype 2.

Sammendrag

Munna monoklonala antikroppar producerades mot Haemophilus pleuropneumoniae bakteriens vanligaste serotyp 2. 11 antikroppsproducerande hybridomkulturer med moderat til stark antigenreaktivitet erhölls. 10 av dessa antikroppar var specifika mot isolerade kapselantigener från H. pleuropneumoniae, serotyp 2, medan en antigen reagerade med kapselantigener från bakterier av alla 8 serotyper. En hybridom som producerade antikroppar med en titer av 1:1000 klonades, och dess antikroppsspecifitet undersöktes ytterligare. Bindningen av denna antikropp (1F3) till hela bakterier, och till kapselextrakt isolerade vid olika temperaturer visar att antikroppen är specifik för en termostabil polysaccaridantigen i cellkapseln av H. pleuropneumoniae, serotyp 2.