Suppr超能文献

开发稳健的冻干工艺以确保 rVSV-SARS-CoV-2 疫苗的长期稳定性。

Development of Robust Freeze-Drying Process for Long-Term Stability of rVSV-SARS-CoV-2 Vaccine.

机构信息

Viral Vectors and Vaccines Bioprocessing Group, Department of Bioengineering, McGill University, Montreal, QC H2X 1Y4, Canada.

出版信息

Viruses. 2024 Jun 11;16(6):942. doi: 10.3390/v16060942.

Abstract

The thermostability of vaccines, particularly enveloped viral vectored vaccines, remains a challenge to their delivery wherever needed. The freeze-drying of viral vectored vaccines is a promising approach but remains challenging due to the water removal process from the outer and inner parts of the virus. In the case of enveloped viruses, freeze-drying induces increased stress on the envelope, which often leads to the inactivation of the virus. In this study, we designed a method to freeze-dry a recombinant vesicular stomatitis virus (VSV) expressing the SARS-CoV-2 spike glycoprotein. Since the envelope of VSV is composed of 50% lipids and 50% protein, the formulation study focused on both the protein and lipid portions of the vector. Formulations were prepared primarily using sucrose, trehalose, and sorbitol as cryoprotectants; mannitol as a lyoprotectant; and histidine as a buffer. Initially, the infectivity of rVSV-SARS-CoV-2 and the cake stability were investigated at different final moisture content levels. High recovery of the infectious viral titer (~0.5 to 1 log loss) was found at 3-6% moisture content, with no deterioration in the freeze-dried cakes. To further minimize infectious viral titer loss, the composition and concentration of the excipients were studied. An increase from 5 to 10% in both the cryoprotectants and lyoprotectant, together with the addition of 0.5% gelatin, resulted in the improved recovery of the infectious virus titer and stable cake formation. Moreover, the secondary drying temperature of the freeze-drying process showed a significant impact on the infectivity of rVSV-SARS-CoV-2. The infectivity of the vector declined drastically when the temperature was raised above 20 °C. Throughout a long-term stability study, formulations containing 10% sugar (sucrose/trehalose), 10% mannitol, 0.5% gelatin, and 10 mM histidine showed satisfactory stability for six months at 2-8 °C. The development of this freeze-drying process and the optimized formulation minimize the need for a costly cold chain distribution system.

摘要

疫苗的热稳定性,尤其是包膜病毒载体疫苗,在需要的任何地方进行递送时仍然是一个挑战。病毒载体疫苗的冷冻干燥是一种很有前途的方法,但由于需要从病毒的内外部分去除水分,因此仍然具有挑战性。在包膜病毒的情况下,冷冻干燥会增加对包膜的压力,这通常会导致病毒失活。在这项研究中,我们设计了一种方法来冷冻干燥表达 SARS-CoV-2 刺突糖蛋白的重组水疱性口炎病毒(VSV)。由于 VSV 的包膜由 50%的脂质和 50%的蛋白质组成,因此制剂研究侧重于载体的蛋白质和脂质部分。制剂主要使用蔗糖、海藻糖和山梨糖醇作为冷冻保护剂;甘露醇作为冻干保护剂;组氨酸作为缓冲剂来制备。最初,在不同最终水分含量水平下,研究了 rVSV-SARS-CoV-2 的感染力和蛋糕稳定性。在 3-6%的水分含量下,发现高回收率的感染性病毒滴度(~0.5 至 1 个对数损失),冻干蛋糕没有恶化。为了进一步降低感染性病毒滴度的损失,研究了赋形剂的组成和浓度。冷冻保护剂和冻干保护剂的浓度从 5%增加到 10%,同时添加 0.5%明胶,导致感染性病毒滴度的回收率提高和稳定的蛋糕形成。此外,冷冻干燥过程的二次干燥温度对 rVSV-SARS-CoV-2 的感染力有显著影响。当温度升高到 20°C 以上时,载体的感染力急剧下降。在长期稳定性研究中,含有 10%糖(蔗糖/海藻糖)、10%甘露醇、0.5%明胶和 10 mM 组氨酸的配方在 2-8°C 下六个月表现出令人满意的稳定性。这种冷冻干燥工艺和优化配方的开发最大限度地减少了对昂贵冷链分配系统的需求。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6a16/11209311/5fa544f3d7ce/viruses-16-00942-g001.jpg

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍。

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索

文档翻译

学术文献翻译模型,支持多种主流文档格式。

立即体验