IMMUNOCHEMICAL ANALYSIS OF INSECT CARBOHYDRATE ANTIGENIC DETERMINANTS- RECOGNITION OF A TERMINAL ALPHA-LINKED N-ACETYLGALACTOSAMINE-CONTAINING EPITOPE OF CALLIPHORA-VICINA NEUTRAL GLYCO(SPHINGO)LIPIDS AND DETECTION IN ADDITIONAL ORTHOPTERAN AND DIPTERAN SPECIES

A cloned mouse hybridoma was established that secreted a monoclonal an tibody directed against certain neutral glycosphingolipids of the thir d-instar larvae of Calliphora vicina (Insecta: Diptera). The isotype o f the designated monoclonal antibody CNF-I (Calliphora Neutral Fractio n) was determined as IgG3. By the use of purified neutral glycosphingo lipids of C.vicina, qualitative thin-layer chromatography immunostaini ng and semi-quantitative enzyme-linked immunosorbent assay determinati ons, the epitope was specified as a terminal alpha1-4-linked N-acetylg alactosamine to subterminal N-acetylgalactosamine, which is present on the components NAcalpha4GalNAcbeta4GlcNAcbeta3Manbeta4Glcbeta1Cer (N5 a) and a4GalNAcbeta4-(6'PEtn-)GlcNAcbeta3Manbeta4GlcB1Cer (Nz5a). Afte r enzymatic removal of the terminal alpha-N-acetylgalactosamine residu e, the epitope reactivity was destroyed. The distribution of the CNF-I recognized epitope among neutral glycolipids of other insects was sho wn for: Locusta migratoria (Insecta: Orthoptera) ; and, Chironomus ten tans and Drosophila melanogaster (Insecta: Diptera).