Plasmonic Inactivation of Virus and Mycoplasma Contaminants

摘要 生物药品，或称“生物制品”，是研发中最重要的药品之一。 今天，它们的安全制造对人类健康绝对至关重要。运营商面临的主要问题 无论是在实验室规模还是在工业规模，微生物污染都是生物反应器的一个整体风险 任何源于活细胞系的过程。根本的问题是确保受污染的生物制品 不会注射到人体内。为了保证无污染的生物制品，通常要进行终端灭菌 这是必要的。在这一步骤中的中心挑战是灭活或去除微生物污染物，而不是 对珍贵的生物制品造成伤害。这项工作的重点是抗体作为代表性的生物制品。特别是 对于病毒和支原体污染，最终的灭菌步骤仍然具有挑战性，因为 病原体的大小。今天的行业标准是通过使用滤膜的被动过滤来去除 具有比病毒颗粒更小或相同大小的孔。然而，这种方法需要很长的时间 与高成本相关的处理时间。此外，超滤可以诱导抗体自结合。 并且与新兴的灵活、小规模、护理点生物制品制造技术不兼容。这个 对新的选择性微生物灭活策略的需求也不仅限于生物制品制造领域。这个 Covid19大流行最近说明了有必要采取可靠的病毒灭活策略，有选择性地采取行动 对组织中的病毒，而不是对蛋白质，例如，允许对感染样本进行免疫学分析 在高密封性实验室之外。光具有杀菌性能，而紫外光长期以来一直被用于 灭活大范围的微生物病原体。不幸的是，它缺乏特异性，也损害了宝贵的 由分子在紫外光范围内的吸收驱动的反应光化学反应的生物制品 电磁频谱。克服超滤和紫外线照射作为微生物的缺点 灭活策略，这项建议开发了一种等离子体增强的光子灭活方法，该方法 利用近红外(NIR)光选择性灭活病毒和支原体。就像近红外辐射一样 与分子吸收不重叠，对生物制品的附带损害最小。拟议的工作将 揭示血浆病原体灭活的基本工作原理并实施磁力灭活 等离子体纳米粒子(NPs)允许轻松、非接触地将纳米材料从 灭菌后的样品。本申请的具体目标是： 目标1：通过等离子体增强实现近红外光可靠的病毒灭活 目的2：用近红外光演示一种等离子体增强支原体灭活策略 目的3：展示磁性等离子体纳米粒对病毒和支原体的可扩展清除

SUMMARY Biological pharmaceuticals, or “biologics”, are among the most important pharmaceuticals in development today, and their safe manufacture is absolutely crucial for human health. A major problem faced by operators of bioreactors, at both the laboratory scale and industrial scale, is microbial contamination as an integral risk of any process that derives from live cell lines. The fundamental concern is to ensure that contaminated biologics are not injected into the human body. To warrant contamination free biologics a terminal sterilization is often necessary. The central challenge in this step is the inactivation or removal of microbial contaminates without causing harm to the precious biologics. This work focuses on antibodies as representative biologics. Especially for viral and mycoplasma contaminations the terminal sterilization step remains challenging due to the small size of the pathogens. The industry standard today is removal through passive filtration using filter membranes with pore diameters smaller or of the same size as the virus particles. This approach requires, however, long processing times associated with high costs. Furthermore, ultrafiltration can induce antibody self-association and is not compatible with emerging flexible, small-scale, point-of-care biologics fabrication technologies. The need for new selective microbe inactivation strategies is also not limited to the field of biologics fabrication. The Covid19 pandemic has recently illustrated the need for reliable virus inactivation strategies that selectively act on the virus in tissues but not on proteins, for instance, to allow immunological assays of infected samples outside of high containment laboratories. Light has sterilization properties, and UV-light has long been used to inactivate a broad range of microbial pathogens. Unfortunately, it lacks specificity and also damages precious biologics through reactive photochemistries driven by molecular absorptions in the UV range of the electromagnetic spectrum. To overcome the shortcomings of both ultrafiltration and UV-irradiation as microbe inactivation strategies, this proposal develops a plasmonically enhanced photonic inactivation method that utilizes near-infrared (NIR) light for the selective inactivation of viruses and mycoplasma. As NIR radiation does not overlap with molecular absorptions, the collateral damage on biologics is minimal. The proposed work will reveal the fundamental working principles underlying plasmonic pathogen inactivation and implement magnetic plasmonic nanoparticles (NPs) that allow for an easy, contact-free removal of the nanomaterials from the samples after sterilization. The specific aims of this application are to: Aim 1: Achieve Reliable Virus Inactivation with NIR Light through Plasmonic Enhancement Aim 2: Demonstrate a Plasmon-Enhancement Strategy for Mycoplasma Inactivation with NIR Light Aim 3: Demonstrate Scalable Clearance of Virus and Mycoplasma with Magnetic Plasmonic NPs