Purification and immunochemical characterization of a 22 kilodalton surface antigen from Schistosoma mansoni

• Published in: Molecular and biochemical parasitology Vol. 28; no. 2; pp. 95 - 103
• Main Authors: Oligino, Lynette D., Percy, Amy J., Harn, Donald A.
• Format: Journal Article
• Language: English
• Published: Shannon Elsevier B.V 01.03.1988; Elsevier Science
• Subjects: Animals; Antigen purification; Antigens, Helminth - immunology; Antigens, Helminth - isolation & purification; Antigens, Surface - immunology; Antigens, Surface - isolation & purification; Biological and medical sciences; Chromatography, Affinity; Electrophoresis, Polyacrylamide Gel; Epitope characterization; Epitopes - immunology; Epitopes - isolation & purification; Fluorescent Antibody Technique; Human protozoal diseases; Immunoassay; Infectious diseases; Irradiated cercariae; Malaria; Medical sciences; Parasitic diseases; Protein Biosynthesis; Protozoal diseases; RNA - genetics; Schistosoma mansoni; Schistosoma mansoni - genetics; Schistosoma mansoni - immunology; Surface membrane; Tropical medicine; PBS; Schistosoma mansoni; Epitope characterization; SDS; Surface membrane; MS; DOC; 2-D; Antigen purification; IrCS; Irradiated cercariae; Purification; Antigenic determinant; Plathelmintha; Identification; Immunochemistry; Cell surface; Molecular weight; Trematoda; Antigen; Cercaria; Animal; Helmintha; Irradiation; Invertebrata
• ISSN: 0166-6851; 1872-9428
• DOI: 10.1016/0166-6851(88)90056-4

A 22 kDa antigen (Sm22) was purified from schistosomula membrane extracts by immunoaffinity chromatography with monoclonal antibody M.2. Western blotting suggested that the epitope bound by M.2 required a specific conformational folding of the molecule, which was sensitive to reducing agents. Two-dimensional electrophoresis of purified Sm22 demonstrated that the single 22 kDa protein recognized by M.2 on one-dimensional gel analysis was composed of at least two isomorphs. Additional Western blotting showed that Sm2 was one of the major antigens recognized by mouse anti-irradiated cercariac serum, and that this same serum recognized at least one epitope which was not sensitive to reducing agents. The mice vaccinated with irradiated cercariae were shown to be 75% protected from cercarial challenge. Sera from a rabbit immunized with Sm22 contained antibodies which bound to the surface of schistosomula and detected a single protein at 22 kDa by immunoprecipitation or Western blot. The rabbit anti-Sm22 sera also immunoprecipitated a 22 kDa in vitro translation product, indicating that at least one epitope on Sm22 is not dependent on glycosylation.