Vessely Christina, Estey Tia, Randolph Theodore W, Henderson Ian, Cooper Julianne, Nayar Rajiv, Braun Latoya Jones, Carpenter John F
Department of Pharmaceutical Sciences. School of Pharmacy, SOP-215, Campus Box C238, University of Colorado Health Sciences Center, Denver, Colorado 80262, USA.
J Pharm Sci. 2009 Sep;98(9):2970-93. doi: 10.1002/jps.21498.
The adsorption of recombinant botulinum neurotoxin (BoNT) protein-derived vaccine antigens to aluminum salt adjuvants has been previously studied for the development of a trivalent vaccine against the neurotoxins (Vessely et al., in press, J Pharm Sci). The current paper describes an investigation of the stability of recombinant BoNT antigens adsorbed to aluminum salt adjuvants during storage in aqueous solution. Both chemical and physical changes occurred during storage. Phosphate groups in the buffer exchanged with hydroxyl groups on the adjuvant surface. The resulting changes in solution pH and adjuvant surface chemistry promoted more favorable electrostatic interaction between the BoNT proteins and the surface, possibly increasing the affinity of the proteins for the surface during storage. Fluorescence and UV spectroscopy suggested changes to protein structure during storage, whereas differential scanning calorimetry showed changes to thermal processes related to protein conformation and/or surface adsorption. The consequence of the chemical and physical changes to the proteins was a decrease in the ability to desorb protein from the adjuvant surface during storage. Overall, the results of this study emphasize the utility of a thorough characterization of the interactions between protein antigens and aluminum salt adjuvants.
此前，为研发一种针对肉毒杆菌神经毒素的三价疫苗，已对重组肉毒杆菌神经毒素（BoNT）蛋白衍生的疫苗抗原与铝盐佐剂的吸附情况展开研究（Vessely等人，即将发表于《药物科学杂志》）。本文描述了对吸附于铝盐佐剂的重组BoNT抗原在水溶液储存期间稳定性的研究。储存过程中发生了化学和物理变化。缓冲液中的磷酸基团与佐剂表面的羟基发生交换。溶液pH值和佐剂表面化学性质的变化促进了BoNT蛋白与表面之间更有利的静电相互作用，这可能会增加储存期间蛋白对表面的亲和力。荧光光谱和紫外光谱表明储存期间蛋白质结构发生了变化，而差示扫描量热法显示与蛋白质构象和/或表面吸附相关的热过程发生了变化。蛋白质化学和物理变化的结果是储存期间从佐剂表面解吸蛋白质的能力下降。总体而言，本研究结果强调了全面表征蛋白质抗原与铝盐佐剂之间相互作用的实用性。
