A novel method for the rational construction of well-defined immunogens: the use of oximation to conjugate cholera toxin B subunit to a peptide-polyoxime complex.
Cholera toxin B subunit (CTB), capable of binding to all mucous membranes in its pentameric form, is a potential carrier of mucosal vaccines. In our previous work we reported that the N-terminus of CTB, a threonine, could in principle undergo oxidation and oximation to form conjugates with a cascade of immunogenic peptides. In this study, we set up a model by chemically coupling CTB to a polyoxime that possessed five copies of influenza virus-derived peptides displayed in comblike form. The construct was reconstituted into pentameric form when eluted from a Superdex column after conjugation, and the pentameric nature of this CTB-viral peptide complex was confirmed by SDS-PAGE. GM(1)-ELISA assay showed that the binding properties of CTB-viral peptide complex were increased 4-5-fold over native CTB.
Chen
Jianhua
J
Department of Medical Biochemistry, University Medical Center, rue Michel-Servet 1, 1211 Geneva 4, Switzerland.
Zeng
Weiguang
W
Offord
Robin
R
Rose
Keith
K
D002772:Cholera Toxin / Q000138:chemical synthesis* / Q000276:immunology* / Q000378:metabolism; D022121:Cholera Vaccines / Q000138:chemical synthesis* / Q000276:immunology* / Q000378:metabolism; D010091:Oximes / Q000138:chemical synthesis* / Q000378:metabolism; D021122:Protein Subunits / Q000138:chemical synthesis* / Q000276:immunology* / Q000378:metabolism; D022223:Vaccines, Subunit / Q000138:chemical synthesis* / Q000276:immunology / Q000378:metabolism; D014614:Vaccines, Synthetic / Q000737:chemistry / Q000276:immunology / Q000378:metabolism
D022121:Cholera Vaccines; D010091:Oximes; D021122:Protein Subunits; D022223:Vaccines, Subunit; D014614:Vaccines, Synthetic; D002772:Cholera Toxin