PMID: 7541431Jun 28, 1995Paper

Enzymatically mediated, glycosidic conjugation of immunoglobulins with viral epitopes

Journal of Immunological Methods
T D BrumeanuC A Bona

Abstract

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

References

Feb 1, 1978·European Journal of Immunology·G KöhlerM J Shulman
Jun 1, 1991·Proceedings of the National Academy of Sciences of the United States of America·R BillettaM Zanetti
Apr 1, 1991·Proceedings of the National Academy of Sciences of the United States of America·C V HardingE R Unanue
Jan 1, 1990·Bioconjugate Chemistry·G E Means, R E Feeney
Jun 1, 1985·Archives of Biochemistry and Biophysics·G Avigad
Jan 1, 1989·Methods in Cell Biology·A L TarentinoT H Plummer
Feb 1, 1987·International Journal of Peptide and Protein Research·R E Feeney
Aug 1, 1982·Analytical Biochemistry·J C CabacunganR E Feeney
Mar 1, 1981·International Journal of Peptide and Protein Research·K F GeogheganR E Feeney
Dec 1, 1994·Journal of Pharmaceutical Sciences·M WeitzhandlerN Avdalovic
Mar 15, 1993·Journal of Immunological Methods·T D BrumeanuH Zaghouani

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Citations

Jun 1, 1996·Immunotechnology : an International Journal of Immunological Engineering·T D BrumeanuH Zaghouani
Mar 14, 2002·International Reviews of Immunology·S CasaresT D Brumeanu

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