Production and characterization of monoclonal antibodies against Actinobacillus pleuropneumoniae serotype 1.

Three monoclonal antibodies (mAbs) against Actinobacillus pleuropneumoniae serotype 1, designated 4.2 A11 B5, 5.1 G8 F10 and 1.5 C5 F4 (IgG3, IgG2b and IgM respectively), were produced and characterized. mAbs 4.2 A11 B5 and 5.1 G8 F10 were directed against different epitopes located in the O chain of the LPS. Both clones also recognized reference strains of A. pleuropneumoniae serotypes 9 and 11. The mAb 1.5 C5 F4 reacted with the reference strain of A. pleuropneumoniae serotype 1, with the encapsulated strain 4045 (but not with its non-capsulated mutant) and with A. pleuropneumoniae serotype 1 purified capsular polysaccharides (CPS). The epitope was sensitive to periodate oxidation, heat-labile, and located in the capsular material of A. pleuropneumoniae serotype 1, as demonstrated by immunoblotting. Treatment of the CPS with 5% ammonium hydroxide eliminated the reaction, which may indicate that the epitope recognized by 1.5 C5 F4 mAb is a O-acetyl containing determinant. When different A. pleuropneumoniae field strains were tested, the percentage of strains recognized by the mAbs varied with the mAb and the test used. Cross-reactions associated with the LPS of some A. pleuropneumoniae serotype 5 field strains could be observed with the 4.2 A11 B5 mAb. Of the three mAbs characterized, 1.5 C5 F4 seemed to be the most suitable for A. pleuropneumoniae serotype 1 detection since it reacted with 99% of serotype 1 field strains and it did not recognize any of the strains belonging to other serotypes.