title: Anthrax sub-unit vaccine: the structural consequences of binding rPA83 to Alhydrogel®.
abstract: An anthrax sub-unit vaccine, comprising recombinant Protective Antigen (rPA83) and aluminium hydroxide adjuvant (Alhydrogel®) is currently being developed. Here, a series of biophysical techniques have been applied to free and adjuvant bound antigen. Limited proteolysis and fluorescence identified no changes in rPA83 tertiary structure following binding to Alhydrogel and the bound rPA83 retained two structurally important calcium ions. For adsorbed rPA83, differential scanning calorimetry revealed a small reduction in unfolding temperature but a large decrease in unfolding enthalpy whilst urea unfolding demonstrated unchanged stability but a loss of co-operativity. Overall, these results demonstrate that interactions between rPA83 and Alhydrogel have a minimal effect on the folded protein structure and suggest that antigen destabilisation is not a primary mechanism of Alhydrogel adjuvancy. This study also shows that informative structural characterisation is possible for adjuvant bound sub-unit vaccines.
authors.affiliation: Institute for Cell and Molecular Sciences, Newcastle University, Newcastle-upon-Tyne, UK.
authors.forename: Andrei
authors.initials: A
authors.lastname: Soliakov
authors.forename: Ian F
authors.initials: IF
authors.lastname: Kelly
authors.forename: Jeremy H
authors.initials: JH
authors.lastname: Lakey
authors.forename: Allan
authors.initials: A
authors.lastname: Watkinson
mesh_terms: D000276:Adjuvants, Immunologic / Q000737:chemistry*; D000327:Adsorption; D000536:Aluminum Hydroxide / Q000737:chemistry*; D022122:Anthrax Vaccines / Q000737:chemistry*; D002118:Calcium / Q000737:chemistry; D002152:Calorimetry, Differential Scanning / Q000379:methods; D016014:Linear Models; D008958:Models, Molecular; D020011:Protective Agents / Q000737:chemistry*; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D058767:Protein Unfolding; D059748:Proteolysis; D011994:Recombinant Proteins / Q000737:chemistry*; D013696:Temperature; D013816:Thermodynamics; D014508:Urea / Q000737:chemistry
chemical_list: D000276:Adjuvants, Immunologic; D022122:Anthrax Vaccines; D020011:Protective Agents; D011994:Recombinant Proteins; D000536:Aluminum Hydroxide; D014508:Urea; D002118:Calcium