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

基于有机离子的新型技术，可在环境温度下长期保存病毒

• 批准号: 10662945
• 负责人: Scott F Michael
• 金额: $ 15.5万
• 依托单位: COLLEGE AT OSWEGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10662945
• 关键词: Accounting; Acids; Affect; Ammonium; Anions; Bacteriophages; Base Pairing; Benign; Binding; Biological Assay; Buffers; COVID-19; COVID-19 vaccine; Capsid Proteins; Carbon; Cations; Characteristics; Circular Dichroism Spectroscopy; Cold Chains; Cryopreservation; DNA; Dengue Virus; Developing Countries; Development; Differential Scanning Calorimetry; Effectiveness; Electrostatics; Enzymes; Exhibits; Family; Fluorides; Formulation; Freezing; Future; Goals; Hydration status; Hydrogen Bonding; Hydrolysis; Ions; Knowledge; Libraries; Life; Lipid Bilayers; Lipids; Liquid substance; Logistics; Lytic; Maintenance; Medicine; Messenger RNA; Methods; Mind; Modeling; Molecular; Monitor; Morphology; Names; Nature; Nucleic Acid Vaccines; Nucleic Acids; Outcome; Polymerase Chain Reaction; Procedures; Process; Production; Property; Proteins; Protocols documentation; Quantitative Structure-Activity Relationship; Recombinant Proteins; Recovery; Resource-limited setting; Resources; Reverse Transcription; Roentgen Rays; Salts; Sampling; Savings; Scanning; Scientist; Solvents; Structure; Structure-Activity Relationship; Techniques; Technology; Temperature; Time; Toxicology; Transmission Electron Microscopy; Transportation; Vaccination; Vaccines; Variant; Viral; Viral Genome; Viral Proteins; Virus; Virus-like particle; Water; aqueous; base; biobank; biomaterial compatibility; cost; cost effective; cytotoxicity; design; fighting; flu; functional group; global health; improved; light scattering; liquid formulation; neglect; novel; novel vaccines; nucleic acid stability; pandemic disease; particle; preservation; prevent; programs; rational design; simulation; solute; stressor; technology development; thermostability; vaccination outcome; vaccine access; vaccine distribution; water diffusion

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

项目夏季 目前大多数可用的疫苗，特别是活疫苗和基于 mRNA 的 COVID-19 疫苗，都是对温度敏感的。 并需要严格的冷链维护，需要在建议的温度下储存和分配 从生产到管理的各个环节，这必然对全球贸易构成最令人望而却步的障碍。 免疫接种计划，特别是在发展中国家，占交付成本的 80%。 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 病毒样本无需冷链或复杂的样本回收方案。 该提案旨在开发一个有机离子平台，用于在常温下长期储存病毒。 面对对新疫苗的需求不断增长，有可能降低成本并避免劳动密集型 与当前生物样本库技术相关的维护 - 离子液体 (IL) – 完全由有机盐组成。 离子 - 提供了一个非常合适的平台，在该平台上可以通过选择离子来改变特性，从而使 我们捕获了用于病毒稳定的溶剂/介质的可调设计。 约 20 wt% 的水可以防止病毒基因组和蛋白质衣壳的水解和酶促降解， 我们将使用肌病毒家族的噬菌体作为保存病毒的可靠方法。 裸露蛋白质颗粒的例子和登革热病毒作为脂质包膜颗粒的例子首先，我们将。 通过杂环阳离子的系统变化开发一个精心设计的新型离子液体库 亲液阴离子，并明智地将两种功能（NH3+和SO2F）纳入IL结构中。 其次，我们将通过配对新类型的生物相容性阳离子和阴离子来实现结构变异。 通过评估其结构完整性、热稳定性和稳定性来检查其稳定病毒的有效性 我们将监测病毒二级结构、热变性的变化。 最后，我们将研究它们的经验结构-活性关系以获得全面的了解。 对病毒之间相互作用的结合特征和分子机制有深入的了解 通过模拟、晶体学和光谱方法研究颗粒和目标水性离子溶剂。 该项目将为病毒在环境温度下长期保存提供可行的解决方案 通过开发对水解稳定的病毒-IL-水基质，可以延长周期（可能长达数十年） 该方法的另一个重要特征是这些核酸-IL。 溶液可以直接通过 PCR 进行扩增，无需事先进行提取、纯化或定量 这种方法的优点是简单，使得过程可以在环境温度下存储和使用。 分布广泛有效，提高了生物样品的长时间稳定性，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。 夏日