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

项目摘要 将超临界二氧化碳和共消毒剂相结合的过程在 条件相对温和。微生物失活的机理尚未得到很好的描述，但破坏 脂质膜的损伤和轻微的氧化损伤似乎与此有关。这一过程，也是高度 高效抗病毒，与其他灭菌方法相比被认为是非常温和的。实验 有证据表明，对蛋白质的损害是有限的，免疫原性表位可以在这个过程中存活下来。在……里面 这个项目NovaSterilis将寻求优化超临界二氧化碳灭菌工艺，以获得广泛和有效的 在试图保持模型蛋白质的完整性的同时进行杀毒治疗。第一个目标是充分 验证超临界过程以对抗病毒作为替代辐射的方法 有效的病毒灭活和损害敏感产品。第二个目标将是描述和描述 优化蛋白质超临界杀菌工艺。蛋白质已经变成了 作为疫苗、治疗药物和组合设备越来越受欢迎。然而，他们的绝育手术仍然存在 极具挑战性。与新墨西哥大学的Bryce Chackerian小组合作， 以及加州大学阿默斯特分校的斯蒂芬·艾尔斯博士和伊戈尔·卡尔塔索夫教授，超临界灭菌的效果 关于生长因子和重组疫苗的研究将通过体外和体内功能试验进行，并将 首次使用最先进的质谱仪以极高的精度测定scCO2的影响 单独或与灭菌剂联合使用对模型蛋白质的构象和氧化状态的影响。

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.