Development of Monoclonal Antibodies to the Extracellular Products ofMycobacteriumspp. Isolated from Chevron Snakehead and the Reference StrainMycobacterium chelonei

Abstract

Thirteen monoclonal antibodies (MAbs) were prepared against the extracellular product (ECP) of Mycobacterium sp., strain TB267, isolated from chevron snakehead Channa striata, and five MAbs were prepared against the reference strain Mycobacterium chelonei. The reactivity of the MAbs was examined against seven different strains of mycobacteria by an enzyme-linked immunosorbent assay (ELISA), and Western blot analysis was used to establish the molecular weight of antigens recognized by the MAbs. Western blot analysis proved essential for the selection of the MAbs because some that did not react by ELISA were found to be positive in this assay. All MAbs recognized a 65-kilodalton (kDa) protein present in ECPs, whole cell sonicates, and lysate preparations of the mycobacteria examined, and the epitopes recognized by the MAbs were located on molecules susceptible to protease activity. The 65-kDa protein, one of the major protein constituents of the mycobacterial preparations, was found in periplasmic spaces and cell walls of the bacteria via immunogold staining with MAbs.