Effect of pH on stability of recombinant botulinum serotype A vaccine in aqueous solution and during storage of freeze-dried formulations.
Effect of pH on stability of recombinant botulinum serotype A vaccine in aqueous solution and during storage of freeze-dried formulations.
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DOI：
10.1002/jps.21409
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发表时间：
2008-12
影响因子：
3.8
通讯作者：
Shouvik Roy;I. Henderson;R. Nayar;T. Randolph;J. Carpenter
中科院分区：
文献类型：
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作者：
Shouvik Roy;I. Henderson;R. Nayar;T. Randolph;J. Carpenter
本研究的目的是评价冻干前溶液pH值对A型肉毒神经毒素（rBoNTA（H（c）稳定性的重要性。这种理解对于蛋白质如rBoNTA（H（c））（多价疫苗产品的潜在成分）具有重要意义。对于多价疫苗，可能难以鉴定满足所有组成蛋白抗原的稳定性要求的液体制剂。因此，冻干多价疫苗制剂可能是更可行的替代方案。因此，评价冻干前pH（可能是次优的）对抗原如rBoNTA（H（c））在冻干/储存期间的稳定性的影响变得重要。我们假设当rBoNTA（H（c））从次优pH冻干时，使用适当的稳定剂可以在冻干和长期储存期间提供足够的物理化学稳定性。我们确定了pH 5和8，其中蛋白质是稳定的和不稳定的聚集。鉴定了可在冻干和储存期间在pH 5的稳定溶液中稳定rBoNTA（H（c））的赋形剂。这些辅料为7.5%（w/v）海藻糖和2.5%（w/v）海藻糖与2.5%（w/v）羟乙基淀粉，含和不含0.01%（w/v）聚山梨酯20。为了支持我们的假设，发现这些赋形剂在冻干/储存期间，当从pH 8的预冻干溶液冻干时，为rBoNTA（H（c））提供足够的物理化学稳定性。
The purpose of this study was to evaluate the importance of prelyophilization solution pH on the stability of botulinum neurotoxin, serotype A (rBoNTA(H(c))). This understanding is of significant importance for proteins such as rBoNTA(H(c)), a potential constituent of a multivalent vaccine product. For multivalent vaccines it may be difficult to identify a liquid formulation satisfying the stability requirements for all constituent protein antigens. Consequently, a lyophilized multivalent vaccine formulation may be a more viable alternative. Therefore evaluating the effect of prelyophilization pH (may be suboptimal) on the stability of antigens such as rBoNTA(H(c)) during lyophilization/storage becomes important. We hypothesize that when rBoNTA(H(c)) is lyophilized from a suboptimal pH, using the appropriate stabilizers can provide adequate physicochemical stability during lyophilization and long-term storage. We identified pH 5 and 8 in which the protein was stable and unstable against aggregation. Excipients were identified that could stabilize rBoNTA(H(c)) during lyophilization and storage in a stable solution of pH 5. These excipients were 7.5% (w/v) trehalose and 2.5% (w/v) trehalose with 2.5% (w/v) HES, with and without 0.01% (w/v) polysorbate 20. In support of our hypothesis, these excipients were found to provide adequate physicochemical stability to rBoNTA(H(c)) during lyophilization/storage, when freeze-dried from a prelyophilized solution of pH 8.