Vessely Christina, Estey Tia, Randolph Theodore W, Henderson Ian, Nayar Rajiv, Carpenter John F
Department of Pharmaceutical Sciences, School of Pharmacy, University of Colorado Health Sciences Center, Denver, Colorado, USA.
J Pharm Sci. 2007 Sep;96(9):2375-89. doi: 10.1002/jps.20880.
Botulinum neurotoxin (BoNT) is a biological warfare threat. Protein antigens have been developed against the seven major BoNT serotypes for the development of a recombinant protein vaccine. This study is an evaluation of adsorption profiles for three of the recombinant protein antigens to aluminum salt adjuvants in the development of a trivalent vaccine against BoNT. Adsorption profiles were obtained over a range of protein concentrations. The results document that charge-charge interactions dominate the adsorption of antigen to adjuvant. Optimal conditions for adsorption were determined. However, potency studies and solution stability studies indicated the necessity of using aluminum hydroxide adjuvant at low pH. To improve the adsorption profiles to AlOH adjuvant, phosphate ions were introduced into the adsorption buffers. The resulting change in the adjuvant chemistry led to an improvement of adsorption of the BoNT antigens to aluminum hydroxide adjuvant while maintaining potency. Competitive adsorption profiles were also determined, and showed changes in maximum adsorption from mixed solutions compared to adsorption from individual protein solutions. The adsorption profiles for each protein varied due to differences in adsorption mechanism and affinity for the adjuvant surface. These results emphasize the importance of evaluating competitive adsorption in the development of multivalent vaccine products.
肉毒杆菌神经毒素（BoNT）是一种生物战威胁。针对七种主要的BoNT血清型已开发出蛋白质抗原，用于重组蛋白疫苗的研发。本研究评估了三种重组蛋白抗原在三价BoNT疫苗研发中对铝盐佐剂的吸附情况。在一系列蛋白质浓度范围内获得了吸附情况。结果表明，电荷相互作用在抗原与佐剂的吸附中起主导作用。确定了最佳吸附条件。然而，效力研究和溶液稳定性研究表明，在低pH值下使用氢氧化铝佐剂是必要的。为改善对AlOH佐剂的吸附情况，在吸附缓冲液中引入了磷酸根离子。佐剂化学性质的这种变化导致BoNT抗原对氢氧化铝佐剂的吸附得到改善，同时保持了效力。还确定了竞争性吸附情况，结果显示与单个蛋白质溶液的吸附相比，混合溶液的最大吸附量发生了变化。由于吸附机制和对佐剂表面亲和力的差异，每种蛋白质的吸附情况各不相同。这些结果强调了在多价疫苗产品研发中评估竞争性吸附的重要性。
