Anthrax sub-unit vaccine: the structural consequences of binding rPA83 to Alhydrogel®.
Anthrax sub-unit vaccine: the structural consequences of binding rPA83 to Alhydrogel®.
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炭疽亚单位疫苗：rPA83 与 AlHydrogel® 结合的结构后果。
DOI：
10.1016/j.ejpb.2011.09.009
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
Watkinson,Allan
中科院分区：
文献类型：
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作者：
Soliakov,Andrei;Kelly,IanF;Lakey,JeremyH;Watkinson,Allan
目前正在开发包含重组保护性抗原（rPA 83）和氢氧化铝佐剂（Alhydrogel®）的炭疽亚单位疫苗。在这里，一系列生物物理技术已被应用于游离和佐剂结合抗原。有限的蛋白水解和荧光检测发现，与Alhydrogel结合后，rPA 83三级结构未发生变化，结合的rPA 83保留了两个结构上重要的钙离子。对于吸附的rPA 83，差示扫描量热法显示，展开温度略有降低，但展开焓大幅下降，而尿素展开表现出不变的稳定性，但损失的协同性。总体而言，这些结果表明rPA 83和Alhydrogel之间的相互作用对折叠蛋白质结构的影响极小，并表明抗原不稳定不是Alhydrogel粘附性的主要机制。该研究还表明，佐剂结合亚单位疫苗可能具有信息性结构表征。
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.