DEVELOPMENT OF MONOCLONAL-ANTIBODIES TO THE EXTRACELLULAR PRODUCTS OFMYCOBACTERIUM SPP ISOLATED FROM CHEVRON SNAKEHEAD AND THE REFERENCE STRAIN MYCOBACTERIUM-CHELONEI

Thirteen monoclonal antibodies (MAbs) were prepared against the extrac ellular product (ECP) of Mycobacterium sp., strain TB267, isolated fro m chevron snakehead Channa striata, and five MAbs were prepared agains t the reference strain Mycobacterium chelonei. The reactivity of the M Abs 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 t he MAbs. Western blot analysis proved essential for the selection of t he MAbs because some that did not react by ELISA were found to be posi tive 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 lo cated on molecules susceptible to protease activity. The 65-kDa protei n, one of the major protein constituents of the mycobacterial preparat ions, was found in periplasmic spaces and cell walls of the bacteria v ia immunogold staining with MAbs.