Unique stabilizing mechanism provided by biocompatible choline-based ionic liquids for inhibiting dissociation of inactivated foot-and-mouth disease virus particles.
Unique stabilizing mechanism provided by biocompatible choline-based ionic liquids for inhibiting dissociation of inactivated foot-and-mouth disease virus particles.
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生物相容性胆碱基离子液体提供独特的稳定机制，用于抑制灭活口蹄疫病毒颗粒的解离
作者：
灭活病毒和病毒样颗粒（VLP）是用于疫苗、免疫治疗和肿瘤治疗的重要类别的生物药物。它们复杂的颗粒结构在加工和储存过程中很容易变性，导致其生物功能丧失。离子液体（IL）作为稳定赋形剂已经在基于蛋白质的药物研究中获得了兴趣，但其对灭活病毒抗原的稳定能力仍然未知。在这里，三种生物相容性胆碱基IL，包括[Cho][H2PO4]，[Cho][Cl]和[Cho][SO4]，被测试作为灭活口蹄疫病毒（iFMDV）的潜在稳定剂，iFMDV是非常不稳定的病毒颗粒，容易解离成更小的五聚体，称为12S。基于差示扫描荧光技术的热稳定性分析，结合高效分子排阻色谱法的146S定量分析，发现[Cho][Cl]和[Cho][SO4]可以提高iFMDV颗粒的热稳定性和长期储存稳定性，而[Cho][H2PO4]则表现出去稳定作用。动物实验表明，iFMDV抗原的免疫原性在所有三种IL中均未减弱。通过监测不同离子液体中病毒颗粒的微环境pH，观察到[Cho][Cl]和[Cho][SO4]中的质子强度比缓冲液和[Cho][H2PO4]中的质子强度相对较低。因此，稳定化机制被认为主要是由于在[Cho][Cl]和[Cho][SO 4]中抑制病毒颗粒的五聚体间界面中的组氨酸残基的质子化，这与针对具有相对简单结构的其他蛋白质报道的机制不同。结果表明，具有适当阴离子的基于胆碱的离子液体是有希望的用于iFMDV或其他疫苗抗原的稳定赋形剂。
Inactivated virus and virus-like particles (VLPs) are important classes of biopharmaceuticals for vaccines, immunotherapy and oncotherapy. Their complex particle structures are easily denatured during processing and storage, leading to loss in their biofunctionality. Ionic liquids (ILs) as stabilizing excipients have garnered interest in protein-based pharmaceutical research, but their stabilizing capacity for inactivated virus antigens remains unknown. Here, three biocompatible choline-based ILs, including [Cho][H2PO4], [Cho][Cl], and [Cho][SO4], were tested as potential stabilizers for the inactivated foot-and-mouth disease virus (iFMDV), which are extremely unstable virus particles easily dissociating into smaller pentamers named 12S. Based on differential scanning fluorimetry technology for thermal stability analysis, together with high-performance size-exclusion chromatography for quantitative determination of 146S, it was found that [Cho][Cl] and [cho][SO4] can improve the thermo- and long-term storage stability of iFMDV particles, while [Cho][H2PO4] showed a destabilizing effect. Animal experiments indicated that the immunogenicity of iFMDV antigens was not attenuated in all three ILs. By monitoring the microenvironmental pH of the virus particles in different ILs, a relatively lower proton intensity was observed in [Cho][Cl] and [Cho][SO4] than in buffers and [Cho][H2PO4]. Therefore, the stabilizing mechanism was supposed to be mainly due to suppression of protonation of histidine residues in the inter-pentamer interface of virus particles in [Cho][Cl] and [Cho][SO4], which is distinct from the mechanism reported for other proteins with relatively simple structures. The results suggest that the choline-based ILs with appropriate anions are promising stabilizing excipients for iFMDV or other vaccine antigens.
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影响因子：
3.8
作者：
Ellard, FM;Drew, J;King, AMQ
通讯作者：
King, AMQ
影响因子：
3.8
作者：
Kissmann, Julian;Joshi, Sangeeta B.;Middaugh, C. Russell
通讯作者：
Middaugh, C. Russell
DOI：
10.1292/jvms.17-0683
发表时间：
2018-04-27
期刊：
The Journal of veterinary medical science
影响因子：
--
作者：
Yamada M;Fukai K;Morioka K;Nishi T;Yamazoe R;Kitano R;Shimada N;Yoshida K;Kanno T;Sakamoto K;Yamakawa M
通讯作者：
Yamakawa M
影响因子：
3.1
作者：
Levitz, SM;Nong, SH;Simons, ER
通讯作者：
Simons, ER
影响因子：
2.7
作者：
Liang, Te;Yang, Decheng;Yu, Li
通讯作者：
Yu, Li