Purification of SARS-CoV-2 Virus-Like Particles (VLPs) Using a Microfluidic Technique for Downstream COVID-19 Vaccine Production

使用微流体技术纯化 SARS-CoV-2 病毒样颗粒 (VLP)，用于下游 COVID-19 疫苗生产

• 批准号: 553518-2020
• 负责人: Poudineh, Mahla
• 金额: $ 3.64万
• 依托单位: University of Waterloo
• 依托单位国家: 加拿大
• 项目类别: Alliance Grants
• 财政年份: 2020
• 资助国家: 加拿大
• 起止时间: 2020-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=703505
• 关键词: Purification; SARS; CoV; Virus; Like

COVID-19 has become a pandemic primarily because there are no existing treatments. Thus, ending this world health crisis requires the rapid development of vaccines and therapeutics. Mediphage Bioceuticals, Inc. (MBI) is producing specific DNA vaccines based on virus-like particles (VLPs). VLPs are multiprotein structures that closely resemble viruses and can elicit strong immune response. VLPs are completely safe and not infectious because they do not contain any viral genetic material. To produce SARS-CoV-2 VLPs, MBI uses insect cell culture where following the VLP formation, cells are lysed for VLPs extraction. MBI urgently requires a technique to rapidly purify the VLPs from cell lysates with high efficiency and throughput while the quality of VLPs is preserved for downstream vaccine production. In this project, MBI and the Poudineh group at the University of Waterloo will develop such a system based on microfluidic technology. Microfluidics enables rapid, cost-effective, and high through target purification. In this project, we combine bead-based assay to capture specifically the VLPs based on their surface proteins and design and fabricate a microfluidic device that can isolate the bead-VLP complex, enabling to retrieve the VLPs. Specifically, the system combines three modules: a mixing device where the VLPs are mixed with beads coated with the capture agent; a separation module which isolates the beads from the rest of contaminating molecules; and a dissociation chamber where the VLPs are recovered. The success of this project addresses the MBI's urgent need to purify SARS-CoV-2 VLPs where the microfluidic system will be integrated into their vaccine production line.

COVID-19已成为一种流行病，主要是因为没有现有的治疗方法。因此，结束这场世界卫生危机需要迅速开发疫苗和治疗方法。 Mediphage Bioceuticals，Inc. (MBI)正在生产基于病毒样颗粒（VLP）的特异性DNA疫苗。VLP是一种多蛋白结构，与病毒非常相似，可以引起强烈的免疫反应。VLP是完全安全的，没有传染性，因为它们不包含任何病毒遗传物质。为了生产SARS-CoV-2 VLP，MBI使用昆虫细胞培养，其中在VLP形成后，裂解细胞用于VLP提取。MBI迫切需要一种技术，以高效率和高通量从细胞裂解物中快速纯化VLP，同时保留VLP的质量用于下游疫苗生产。 在这个项目中，MBI和滑铁卢大学的Poudineh小组将开发一个基于微流体技术的系统。微流体技术能够实现快速、经济、高效的靶向纯化。在这个项目中，我们结合了联合收割机，以基于其表面蛋白特异性捕获VLP，并设计和制造了一个微流控装置，可以分离珠-VLP复合物，使回收的VLP。具体地，该系统结合了三个模块：混合装置，其中VLP与包被有捕获剂的珠粒混合;分离模块，其将珠粒与其余污染分子分离;以及解离室，其中回收VLP。 该项目的成功解决了MBI对纯化SARS-CoV-2 VLP的迫切需求，微流体系统将被集成到他们的疫苗生产线中。