Department of Chemical and Biological Engineering, Center for Pharmaceutical Biotechnology, University of Colorado, Boulder, CO 80309, United States.
Department of Chemical and Biological Engineering, Center for Pharmaceutical Biotechnology, University of Colorado, Boulder, CO 80309, United States; Department of Chemical and Biochemical Engineering, Dongguk University-Seoul, Seoul 04620, Republic of Korea.
Eur J Pharm Biopharm. 2019 Mar;136:213-220. doi: 10.1016/j.ejpb.2019.01.019. Epub 2019 Jan 28.
No United States Food and Drug Administration-licensed vaccines protective against Ebola virus (EBOV) infections are currently available. EBOV vaccine candidates currently in development, as well as most currently licensed vaccines in general, require transport and storage under a continuous cold chain in order to prevent potential decreases in product efficacy. Cold chain requirements are particularly difficult to maintain in developing countries. To improve thermostability and reduce costly cold chain requirements, a subunit protein vaccine against EBOV was formulated as a glassy solid using lyophilization. Formulations of the key antigen, Ebola glycoprotein (EBOV-GP), adjuvanted with microparticulate aluminum hydroxide were prepared in liquid and lyophilized forms, and the vaccines were incubated at 40 °C for 12 weeks. Aggregation and degradation of EBOV-GP were observed in liquid formulations during the 12-week incubation period, whereas changes were minimal in lyophilized formulations. Antibody responses against EBOV-GP following three intramuscular immunizations in BALB/c mice were used to determine vaccine immunogenicity. EBOV-GP formulations were equally immunogenic in liquid and lyophilized forms. After lyophilization and reconstitution, adjuvanted vaccine formulations produced anti-EBOV-GP IgG antibody responses in mice similar to those generated against corresponding adjuvanted liquid vaccine formulations. More importantly, antibody responses in mice injected with reconstituted lyophilized vaccine formulations that had been incubated at 40 °C for 12 weeks prior to injection indicated that vaccine immunogenicity was fully retained after high-temperature storage, showing promise for future vaccine development efforts.
目前尚无获得美国食品和药物管理局(FDA)许可的针对埃博拉病毒(EBOV)感染的疫苗。目前正在开发的 EBOV 疫苗候选物，以及一般而言大多数获得许可的疫苗，都需要在冷链下运输和储存，以防止产品功效潜在降低。冷链要求在发展中国家尤其难以维持。为了提高热稳定性并降低昂贵的冷链要求，使用冷冻干燥法将针对 EBOV 的亚单位蛋白疫苗制成玻璃状固体。用微颗粒状氢氧化铝佐剂配制的关键抗原埃博拉糖蛋白(EBOV-GP)的制剂以液体和冷冻干燥形式制备，并将疫苗在 40°C 下孵育 12 周。在 12 周的孵育期间，在液体制剂中观察到 EBOV-GP 的聚集和降解，而在冷冻干燥制剂中变化最小。在 BALB/c 小鼠中进行三次肌肉内免疫后，针对 EBOV-GP 的抗体反应用于确定疫苗的免疫原性。在液体和冷冻干燥形式中，EBOV-GP 制剂具有相同的免疫原性。冷冻干燥和复溶后，佐剂疫苗制剂在小鼠中产生针对 EBOV-GP 的 IgG 抗体反应，与针对相应佐剂液体疫苗制剂产生的反应相似。更重要的是，在注射前已在 40°C 下孵育 12 周的复溶冷冻干燥疫苗制剂注射的小鼠中的抗体反应表明，疫苗免疫原性在高温储存后得到完全保留，为未来的疫苗开发工作带来了希望。
