Novel organic-ion-based technology for long-term virus preservation at ambient temperature

项目总结 目前可用的大多数疫苗，特别是基于新冠肺炎的活疫苗和基于mRNAs的疫苗，都是温度敏感的。 并需要严格的冷链维护，需要在建议的温度下存储和分发 从生产到管理的转变。这种必要性对全球移民构成了最令人望而却步的障碍-- 市民化计划，特别是在发展中国家，占成本交付的80%。因此， 迫切需要一种技术来提供具有成本效益的长期环境温度存储 无需冷链或复杂的样本回收程序即可获得病毒样本。 这项建议旨在开发一种有机离子平台，用于在常温下长期存储病毒。 面对不断增长的新疫苗需求，潜在地降低成本并避免劳动密集型 与当前的生物库技术相关的维护。离子液体(ILS)有机盐完全由 Of Ions提供了一个非常适合的平台，在该平台上可以通过选择离子来改变属性，从而使 用于稳定病毒的溶剂/介质的可调设计。我们假设建议的ILS的解决方案与 约20wt%的水可以防止病毒基因组和蛋白质衣壳的水解性和酶降解， 提供了一种可靠的方法来保存病毒。我们将使用来自肌病毒家族的噬菌体作为 裸露的蛋白质颗粒和登革热病毒的例子，作为脂质包裹的颗粒的例子。首先，我们将 通过杂环阳离子和杂环阳离子的系统变化，开发一个构思周到的新型ILS文库 渗透压负离子，并明智地将两个官能团(NH3+和SO2F)纳入IL结构。 通过将新的生物相容的阳离子和阴离子配对，将实现结构的可变性。第二，我们将 通过评估它们的结构完整性、热稳定性和 保质期从六个月到四年。我们将监测病毒二级结构的变化，热变性- 和颗粒形态。最后，我们将研究它们的经验结构-活性关系，以获得更全面的 对病毒的结合特征和相互作用的分子机制有深入的了解 通过模拟、结晶学和光谱学的方法，对粒子和目标水基离子溶剂进行了研究。 该项目将为病毒的常温保存提供一种可行的解决方案 通过开发对水解性稳定的病毒IL水基质(可能为数十年) 和酶降解。提出的方法的另一个重要特征是这些核酸-ILS 不经提取、纯化或定量，即可直接用PCR扩增溶液。 提顿。这种方法的优点是简单，使得常温储存和处理过程 极其高效的分配，增加了生物样本的长时间稳定性，减少了操作 成本和碳足迹，并改善病毒和基于病毒的技术的物流。 夏季

PROJECT SUMMERY Most currently available vaccines, especially live and mRNA-based COVID-19 vaccines, are temperature sensi- tive and require stringent cold-chain maintenance, entailing their storage and distribution at recommended tem- peratures from production to administration. This necessity imposes the most prohibitive barrier to global im- munization programs, particularly in developing countries, accounting for up to 80% of the cost delivery. Thus, there is a critical need for a technology to provide cost-effective and long-term ambient temperature storage for viral samples without requiring cold chain or complicated sample recovery protocols. This proposal aims to develop an organic-ion platform for long-term storage of viruses at ambient tem- perature to potentially reduce costs in the face of growing needs for new vaccines and avoid labor-intensive maintenance associated with current biobanking technology. Ionic liquids (ILs)  organic salts comprised entirely of ions  offer a well-suited platform on which the properties can be altered by the selection of ions, enabling the tunable design of solvents/media for virus stabilization. We hypothesize that the solutions of proposed ILs with ca. 20 wt% water may prevent hydrolytic and enzymatic degradation of viral genomes and protein capsids, providing a reliable approach to preserve viruses. We will use a bacteriophage from the myovirus family as an example of a naked protein particle and dengue virus as an example of a lipid-enveloped particle. First, we will develop a thoughtfully conceived library of novel ILs through systematic variations of heterocyclic cations and kosmotropic anions, and judicious incorporation of two functionalities (NH3+ and SO2F) into the IL structures. Structural variability will be achieved by pairing new genre of biocompatible cations and anions. Second, we will examine their effectiveness for stabilizing viruses by evaluating their structural integrity, thermostability, and shelf-life from six months and four year. We will monitor changes in viral secondary structure, thermal denatur- ation, and particle morphology. Last, we will study their empirical structure-activity relationships to gain compre- hensive understanding of binding characteristics and molecular mechanisms of interactions between the viral particles and the targeted aqueous ionic solvents via simulation, crystallographic, and spectroscopic methods. This project will provide a viable solution for ambient temperature preservation of viruses for extended periods (potentially for decades) by developing the virusILwater matrices that are stable towards hydrolytic and enzymatic degradation. Another important feature of the proposed approach is that these nucleic acid-ILs solutions can be directly amplified by PCR without being subjected to prior extraction, purification or quantifica- tion. This approach has the merit of simplicity, which makes the process of ambient temperature storage and distribution profoundly efficient, increases the stability of biosamples for prolonged time, reduces operational costs and carbon footprint, and improves logistics for viruses and virus-based technologies. Summery