Production and Utilization of Monoclonal Antibodies against Pseudomonas cepacia

Abstract

To obtain monoclonal antibodies (MABs) specific to Pseudomonas cepacia (Pc) strain RB 425, the live cells (L) and the fixed cells (F) by glutaraldehyde were injected to mice, respectively. Sixteen and 8 hybridoma lines were established from L-immunized and F-immunized mouse, respectively. Three MABs from L-hybridoma lines and another 3 MABs from F-hybridoma lines were used for the specific detection and analyses of the epitopes on Pc. Although some of the 6 MABs of ascitic fluids showed almost the same titers as rabbit antisera in indirect ELI-SA, they had a limited ability to precipitate and agglutinate the L cells. The reactivity of the 6 MABs was tested with L, F, heated L (HL) and heated F (HF) cells in indirect ELISA. Three out of 6 MABs were heterospecific antibodies that reacted more strongly with HL cells than did with each immunogen. Three types of epitopes were classified by the reactivity of MABs with L and F cells. The specificity of MABs was detected by indirect ELISA. The live cells of 5 Pc strains, 6 pseudomonads species, and other bacterial genera were studied for their antigenicity. Five epitopes were distinguished by the reaction patterns of the MABs, and the specificity of the MABs was different in the 5 strains of Pc. Furthermore, 2 MABs from L-hybridoma lines showed a wide specificity in their reaction with pseudomonads, while the others reacted only with the strains of Pc.