Optimization of Supercritical Carbon Dioxide Based Virus Inactivation, Characterized by Protein Damage and Maintenance of Epitope Integrity in Vaccine Sterilization

基于超临界二氧化碳的病毒灭活优化，其特征是疫苗灭菌中的蛋白质损伤和表位完整性的维持

• 批准号: 9253508
• 负责人: Julien Fey
• 金额: $ 60.81万
• 依托单位: NOVASTERILIS, INC.
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2018-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9253508
• 关键词: Address; Adopted; Affect; Allografting; Amino Acids; Area; Attention; Bacterial Spores; Bacteriophages; Biocompatible Materials; Biological Assay; Carbon Dioxide; Cell Line; Collaborations; Confidential Information; Data; Dehydration; Development; Devices; Epitopes; Ethylene Oxide; Genome; Goals; Growth Factor; HIV; Health; Hepatitis A; Hepatitis C; Human Papilloma Virus Vaccine; Human Papillomavirus; Human Resources; Immunization; In Vitro; Individual; Industry; International; Maintenance; Mammalian Cell; Mass Spectrum Analysis; Medical Device; Membrane Lipids; Methods; Microbe; Modeling; Molecular Conformation; Mus; New Mexico; Organism; Patients; Peracetic Acid; Pharmaceutical Preparations; Phase; Play; Process; Property; Proteins; Radiation; Recombinant Vaccines; Reproduction spores; Research; Residual state; Risk; Role; Safety; Sterility; Sterilization; Stress; Structural Protein; Surface; Suspensions; Technology; Testing; Therapeutic; Time; Tissue Banks; Tissues; Universities; Vaccines; Validation; Viral; Virus; Virus Inactivation; Zika Virus; adduct; base; biophysical properties; bone; carcinogenicity; cost; crosslink; immunogenic; improved; in vivo; innovation; link protein; microbicide; microorganism; occupational hazard; oxidation; oxidative damage; particle; pathogen; pressure; professor; protein structure; therapeutic protein; time use; virucide

Project Summary The process combining supercritical CO2 and co-sterilants produces strong synergistic microbicidal effects in relatively mild conditions. The mechanism of microbe inactivation is not well characterized, but the disruption of lipid membranes and mild oxidative damage appear to be involved. The process, which is also highly efficient against viruses, is considered to be very mild compared to other sterilization methods. Experimental evidence shows that damage to protein is limited, and that immunogenic epitopes can survive the process. In this project NovaSterilis will seek to optimize the supercritical CO2 sterilization process for broad and effective virucidal treatments while attempting to maintain the integrity of model proteins. The first goal is to fully validate the supercritical process against viruses as a method that could replace -radiations which fail at effective virus inactivation and damage sensitive products. The second goal will be to characterize and optimize the supercritical sterilization process for the sterilization of proteins. Proteins have become increasingly popular as vaccines, therapeutics, and in combination devices. Yet, their sterilization remains extremely challenging. In collaboration with the group of Bryce Chackerian at the University of New Mexico, and Dr. Stephen Eyles and Professor Igor Kaltashov at UMass-Amherst, the effect of supercritical sterilization on growth factors and recombinant vaccines will be studied with in vitro and in vivo functional assay, and will for the first time use state-of-the-art mass spectrometry to determine with great precision the effect of scCO2 itself or combined with a sterilant on the conformation and oxidation status of model proteins.

项目摘要 结合超临界CO2和辅助灭菌剂的方法产生强的协同杀微生物效果， 相对温和的条件。微生物灭活的机理还没有很好的表征，但破坏 脂质膜和轻度氧化损伤似乎参与其中。这个过程，也是高度 有效的抗病毒，被认为是非常温和的相比，其他灭菌方法。实验 有证据表明，对蛋白质的损伤是有限的，并且免疫原性表位可以在该过程中存活。在 该项目NovaSterilis将寻求优化超临界CO2灭菌工艺， 杀病毒处理，同时试图保持模型蛋白的完整性。第一个目标是全面 验证超临界过程对病毒的作用，作为一种方法，可以取代超临界辐射， 有效的病毒灭活和破坏敏感产品。第二个目标是描述和 优化蛋白质超临界杀菌工艺。蛋白质已经成为 作为疫苗、治疗剂和组合装置越来越受欢迎。然而，他们的绝育仍然存在， 极具挑战性。与新墨西哥州大学的布莱斯·查克里安小组合作， 麻省大学阿默斯特分校的斯蒂芬·埃尔斯博士和伊戈尔·卡尔塔绍夫教授， 将通过体外和体内功能测定研究生长因子和重组疫苗， 第一次使用最先进的质谱法以高精度确定scCO2的影响 其本身或与灭菌剂组合对模型蛋白质的构象和氧化状态的影响。