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This chapter details the characterization of anti-Entamoeba histolytica Monoclonal Antibodies (McAb) that will be restricted to three methods: indirect fluorescent antibody (IFA)-staining patterns, agglutination with live amebic trophozoites, and inhibition of in vitro adherence of amebic trophozoites to target cells. Conventional characterization of anti-E. histolytica McAb involves identification of isotypes and subisotypes of immunoglobulins, identification of molecules recognized by McAb by using immunoprecipitation or Western blot analysis, and determination of their functional activities. Biomedical applications of McAb include typing of pathogenic zymodemes, detection of E. histolytica antigens in clinical specimens, purification of antigenic molecules relevant to vaccine development. McAb-based enzyme-linked immunosorbent assay for detection of E. histolytica can potentially be applied to clinical diagnosis of amebiasis especially when large numbers of specimens are to be processed. Purification of amebic antigens of functional interest can be achieved by affinity chromatography. |
