CHARACTERIZATION OF MONOCLONAL-ANTIBODIES TO THE 26-KDA GLUTATHIONE-S-TRANSFERASE OF SCHISTOSOMA-JAPONICUM

Six monoclonal antibodies (MAbs) were raised in mice against the 26-kD a glutathione S-transferase (GST) of the parasite Schistosoma japonicu m. These MAbs were originally selected for their specific binding to t he recombinant GST (r-GST) generated in E, coli by an enzyme-linked im munosorbent assay, A further study demonstrated that all these MAbs bo und to plate-coated GST affinity-purified from the parasite Schistosom a japonicum, However, in Western blotting analysis only a single monoc lonal antibody (MAb Y3D7) yielded positive binding, The binding of MAb Y3D7 on Western blotting was further characterized; specific binding was found on other GST fusion proteins and on the authentic 26-kDa GST but not the 28-kDa GST in the total soluble worm proteins from Schist osoma japonicum, Using protein-A-mediated immunoprecipitation, MAbs Y3 D7 and Y5D5 precipitated r-GST while in parallel experiments the remai ning MAbs did not generate r-GST precipitation, In an alternative co-p recipitation experiment, r-GST was first bound to glutathione (GSH) Se pharose beads and subsequently tested for interaction with the MAbs, T n this manner, all MAbs except MAb Y5D5 were co-precipitated with the complexes, Thus, these select MAbs readily reacted with GST although t heir binding characteristics were different, Because GST has been wide ly used in the generation of fusion proteins for various purposes and is a potential vaccine candidate in controlling schistosomiasis, these MAbs should prove valuable for their application to molecular biology and parasitology.