Biochemical characterization and localization of Fasciola hepatica 26-28 kDa diagnostic coproantigen.
We have previously reported the usefulness of a 26-28 kDa coproantigen of Fasciola hepatica for diagnosis of infection. In this study, the 26-28 kDa coproantigen was biochemically characterized with the aid of monoclonal antibodies (MoAb) in an effort to better understand the biology of the antigen. Differential staining of chromatographically-purified 26-28 kDa coproantigen on SDS-PAGE, under reducing and non-reducing conditions, indicated that the coproantigen was a monomeric, highly glycosylated glycoprotein. Alkaline treatment of the purified coproantigen resulted in an 8 kDa protein core which still contained the epitope recognized by the MoAb. No protease activity was associated with the 26-28 kDa coproantigen. The coproantigen could be cleaved by trypsin without altering the reactive epitope recognized by the MoAb, but was resistant to pepsin digestion. Further, the coproantigen was stable under several different storage conditions. Indirect immunofluorescence on tissue sections of adult flukes indicated that the coproantigen was present in gut cells and tegument. Taken together these results confirm the stability of the 26-28 kDa coproantigen and its usefulness in diagnostic tests for F. hepatica infections.
Abdel-Rahman
S
S
Department of Veterinary Microbiology and Parasitology, School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA 70803, USA.
O'Reilly
K L
KL
Malone
J B
JB
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000947:Antigens, Helminth / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification*; D015153:Blotting, Western; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis*; D005210:Fasciola hepatica / Q000254:growth & development / Q000276:immunology*; D005211:Fascioliasis / Q000175:diagnosis / Q000276:immunology / Q000662:veterinary*; D019084:Fluorescent Antibody Technique, Indirect; D015801:Helminth Proteins / Q000737:chemistry / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000947:Antigens, Helminth; D015801:Helminth Proteins