Tuckey, Corinna Dawn2024-08-152024-08-1519961996https://hdl.handle.net/1828/19945Six monoclonal antibodies previously derived against internal Trypanosoma brucei rhodesiense ViTat 1.1 procyclic culture form antigens were characterized for species- and life cycle stage-specificity by immunoblotting. Five of the mAbs were specific for T. brucei spp. while the sixth (mAb #20) recognized an antigen in all trypanosome species and life cycle stages tested. Immunogold electron microscopy revealed that mAb #20 immunoreactivity localized in the mitochondrion. The antigen recognized by mAb #20 was partially purified and the N-terminal sequence showed significant identity with T. cruzi Hsp 60 and E. coli GroEL mitochondrial chaperones. Using a combination of expression library screening and polymerase chain reaction amplification, I isolated the cDNA encoding the trypanosome antigen. The translated sequence matched sequences of mitochondrial chaperones from several species. Taken together, the results indicate that the antigen recognized by mAb #20 is the T. brucei spp Hsp60 mitochondrial chaperone. The mAb #20 or the Hsp60 protein may be used in a simple immunodiagnostic test for African sleeping sickness.110 pagesAvailable to the World Wide WebThesis