Partial characterization of the epitope on excretory-secretory products of Fasciola hepatica recognized by monoclonal antibody ES78.
This report contains a partial characterization of the epitope recognized by monoclonal antibody (MAb) ES78 produced against excretory-secretory (ES) antigens of Fasciola hepatica. ES78 is currently used for the detection of ES antigens in serum and stool samples of cattle and humans with fasciolosis, using a highly sensitive and specific sandwich enzyme-linked immunosorbent assay (ELISA). The epitope was characterized by periodate oxidation, alkaline borohydride reduction, trichloroacetic acid precipitation, beta-mercaptoethanol treatment, and enzymatic proteolysis. These results, together with those of the 2-site ELISA, lectin immunoassays, and beta-galactosidase digestion, showed that MAb ES78 reacts with a partly protein/partly carbohydrate antigenic determinant that is found on several ES molecules of adult specimens of F. hepatica and contains at least 1 disulfide bond and beta-galactose probably as galactose-beta(1-3)-N-acetylgalactosamine disaccharide.
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Departamento de Parasitología, Instituto de Medicina Tropical Pedro Kouri, Ciudad de la Habana, Cuba.
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Marcet
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D000818:Animals; D000909:Antibodies, Helminth / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000947:Antigens, Helminth / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification; D001894:Borohydrides / Q000494:pharmacology; D002417:Cattle; D002846:Chromatography, Affinity; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D005210:Fasciola hepatica / Q000276:immunology*; D006358:Hot Temperature; D007118:Immunoassay; D037102:Lectins / Q000378:metabolism; D008623:Mercaptoethanol / Q000494:pharmacology; D009439:Neuraminidase / Q000378:metabolism; D010504:Periodic Acid / Q000494:pharmacology; D011402:Pronase / Q000378:metabolism; D014238:Trichloroacetic Acid / Q000494:pharmacology; D001616:beta-Galactosidase / Q000378:metabolism
D000909:Antibodies, Helminth; D000911:Antibodies, Monoclonal; D000947:Antigens, Helminth; D001894:Borohydrides; D000939:Epitopes; D037102:Lectins; D010504:Periodic Acid; D014238:Trichloroacetic Acid; D008623:Mercaptoethanol; C025364:sodium borohydride; C009288:metaperiodate; D009439:Neuraminidase; D001616:beta-Galactosidase; D011402:Pronase