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 antibody directed against certain neutral glycosphingolipids of the third-instar larvae of Calliphora vicina (Insecta: Diptera). The isotype of the designated monoclonal antibody CNF-I (Calliphora Neutral Fraction) was determined as IgG3. By the use of purified neutral glycosphingolipids of C. vicina, qualitative thin-layer chromatography immunostaining and semi-quantitative enzyme-linked immunosorbent assay determinations, the epitope was specified as a terminal alpha 1-4-linked N-acetylgalactosamine to subterminal N-acetylgalactosamine, which is present on the components GalNAc alpha 4GalNAc beta 4GlcNAc beta 3Man beta 4Glc beta lCer (N5a) and GalNAc alpha 4GalNAc beta 4-(6' PEtn-)GlcNAc beta 3Man beta 4Glc beta lCer (Nz5a). After enzymatic removal of the terminal alpha-N-acetylgalactosamine residue, the epitope reactivity was destroyed. The distribution of the CNF-I recognized epitope among neutral glycolipids of other insects was shown for: Locusta migratoria (Insecta: Orthoptera); and, Chironomus tentans and Drosophila melanogaster (Insecta: Diptera).
Keller
M
M
Institut für Physiologische Chemie, Philipps-Universität Marburg, Germany.
Sorgenfrei
B
B
Dennis
R D
RD
Wiegandt
H
H
D000116:Acetylgalactosamine / Q000276:immunology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000032:analysis / Q000276:immunology*; D002460:Cell Line; D004175:Diptera / Q000196:embryology / Q000254:growth & development / Q000276:immunology*; D000939:Epitopes / Q000737:chemistry / Q000187:drug effects / Q000276:immunology*; D005260:Female; D006026:Glycoside Hydrolases / Q000494:pharmacology; D006028:Glycosphingolipids / Q000276:immunology* / Q000302:isolation & purification; D007814:Larva; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009987:Orthoptera / Q000276:immunology; D013045:Species Specificity
D000911:Antibodies, Monoclonal; D002241:Carbohydrates; D000939:Epitopes; D006028:Glycosphingolipids; D006026:Glycoside Hydrolases; D000116:Acetylgalactosamine