Effect of Osmotic Pressure on the Stability of Whole Inactivated Influenza Vaccine for Coating on Microneedles
Effect of Osmotic Pressure on the Stability of Whole Inactivated Influenza Vaccine for Coating on Microneedles
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DOI：
10.1371/journal.pone.0134431
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发表时间：
2015-07-31
期刊：
影响因子：
3.7
通讯作者：
Prausnitz, Mark R.
中科院分区：
文献类型：
--
作者：
Choi, Hyo-Jick;Song, Jae-Min;Prausnitz, Mark R.
被包裹的病毒疫苗可能会被高渗透强度的溶液破坏，例如那些用于在干燥过程中保护疫苗抗原的溶液，这些溶液含有高浓度的糖。因此，我们研究了整个灭活的流感病毒在高渗溶液中的收缩和活性损失，并利用这些发现来改进流感疫苗接种微针贴片的疫苗涂层。利用停流光散射分析，我们发现病毒在暴露于217毫升的高渗透压差时，在5 S内经历了约10%的体积收缩。在这种收缩过程中，病毒被膜具有很低的渗透性(1-6x10(-4)cm S(-1))和很高的阿累尼乌斯活化能(E-a=15.0kcal mol(-1))，表明水分子通过病毒脂膜扩散。在约20 S至2分钟的准稳定状态后，根据双糖的种类和高渗渗透强度的不同，存在病毒收缩的第二阶段。在最高渗透强度下，这导致了起伏的光散射谱，这似乎与病毒包膜的扰动有关，通过体外血凝测量和小鼠体内免疫原性研究确定了病毒活性的丧失。添加羧甲基纤维素有效地防止了疫苗在体外和体内的活性损失，这被认为是由于增加了浓糖溶液的粘度，从而减少了微针包被过程中的渗透应力。这些结果表明，高渗溶液会导致灭活流感病毒整体的双相收缩，从而在高渗透强度下损害疫苗的活性，而在疫苗包衣溶液中添加增粘剂可以防止渗透驱动的损害，从而能够制备用于疫苗接种的稳定的微针包衣制剂。
Enveloped virus vaccines can be damaged by high osmotic strength solutions, such as those used to protect the vaccine antigen during drying, which contain high concentrations of sugars. We therefore studied shrinkage and activity loss of whole inactivated influenza virus in hyperosmotic solutions and used those findings to improve vaccine coating of microneedle patches for influenza vaccination. Using stopped-flow light scattering analysis, we found that the virus underwent an initial shrinkage on the order of 10% by volume within 5 s upon exposure to a hyperosmotic stress difference of 217 milliosmolarity. During this shrinkage, the virus envelope had very low osmotic water permeability (1-6x10(-4) cm s(-1)) and high Arrhenius activation energy (E-a = 15.0 kcal mol(-1)), indicating that the water molecules diffused through the viral lipid membranes. After a quasi-stable state of approximately 20 s to 2 min, depending on the species and hypertonic osmotic strength difference of disaccharides, there was a second phase of viral shrinkage. At the highest osmotic strengths, this led to an undulating light scattering profile that appeared to be related to perturbation of the viral envelope resulting in loss of virus activity, as determined by in vitro hemagglutination measurements and in vivo immunogenicity studies in mice. Addition of carboxymethyl cellulose effectively prevented vaccine activity loss in vitro and in vivo, believed to be due to increasing the viscosity of concentrated sugar solution and thereby reducing osmotic stress during coating of microneedles. These results suggest that hyperosmotic solutions can cause biphasic shrinkage of whole inactivated influenza virus which can damage vaccine activity at high osmotic strength and that addition of a viscosity enhancer to the vaccine coating solution can prevent osmotically driven damage and thereby enable preparation of stable microneedle coating formulations for vaccination.