Enzymatically mediated, glycosidic conjugation of immunoglobulins with viral epitopes.

摘要: Abstract We developed a novel enzymatic procedure to couple peptide the sugar moieties of immunoglobulins (Igs). The synthesis conjugates consists in galactose (Gal) oxidation desialylated Igs followed by covalent attachment peptides with concurrent stabilization Schiff bases upon mild reduction. used this study, corresponds amino acid residues 110–120 hemagglutinin (HA) PR8 A virus and is recognized CD4 T helper cells association I-Ed class II major histocompatibility complex (MHC). degree coupling as determined competitive inhibition radioimmunoassay (IRIA) using FPLC purified was estimated at 11.4 per IgG molecule. Coupling HA110–120 moiety various mouse human confirmed Western blot analysis anti-HA110–120 antibodies. Complete detachment from N-deglycosylation PGNase F indicated defined specificity HA N-linked oligosaccharides Igs. To facilitate quick release into lysosomal compartment antigen processing (APC) we introduced α terminus (HAc110–120), cleavage site for cathepsins (AAAL). immunoglobulin-galactose-HAc110–120 (IGP) were able activate specific hybridoma efficient influenza 40–100-fold higher than synthetic itself.