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

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Abstract
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. GM1-ELISA assay showed that the binding properties of CTB−viral peptide complex were increased 4−5-fold over native CTB.