Immunochemical analysis of a monoclonal antibody recognizing a terminal glucuronic acid-containing epitope of insect acidic glycolipids

Abstract

A cloned, hybridoma cell-line was established that secreted the monoclonal antibody CAF-I following stimulation of the donor Balb/c mouse spleen cells by the total acidic fraction glycolipids of the third-instar larvae of Calliphora vicina (Insecta:Diptera). The monoclonal antibody isotype was IgG3 By qualitative (TLC-immunostaining) and semi-quantitative (enzyme-linked immunosorbent assay) methods, and comparison with the cross-reactivity of known monoclonal antibodies, the epitope was specifically located on the terminal, non-reducing end of the oligosaccharide chain of most of the insect acidic glycolipids. Following isolation of the two main acidic glycolipids of C. vicina larvae (A5c and Az5c), exoglycosidase treatment characterized the terminal disaccharide CAF-I epitope as glucuronic acid bound to subterminal galactose, both in the beta-anomeric configuration: G1cA beta-Ga1 beta-. The immunohistological distribution of this epitope in the dipteran, Drosophila melanogaster, showed its main expression to be in the imaginal discs and brain of the third-instar larva, and the retinula cells of the ommatidial elements of the compound eye retina of the adult female.