Continuous Production of Viral Vectors using membraneless Perfusion Culture of Host Cells

利用宿主细胞无膜灌注培养连续生产病毒载体

• 批准号: 10414312
• 负责人: JONGYOON HAN
• 金额: $ 60万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10414312
• 关键词: Continuous; Production; Viral; Vectors; using

Project Summary Abstract Continuous Bioproduction of Viral Vectors via High Cell Density, Membrane-less Perfusion Culture In this project, we propose to intensify the viral vector production process by the membrane-less perfusion bioreactor system, which was developed by our team at MIT. Based on the inertial microfluidics and fully validated in CHO cell cultivation, this system has many critical advantages over existing perfusion bioreactors, including high product recovery and avoiding membrane fouling, continuous removal of dead cells, cell debris, and cell aggregates, and easy maintenance and automation. As a result, our bioreactors can maintain high cell density reliably even when the cell viability and culture conditions change significantly, ideally suited for controlling viral vector production processes. Teaming with experts in HEK293 and Sf9 based production systems, we will implement long-term, high cell density cultures of host cells and explore and validate several innovative vector production platforms that provide more accurate control over fresh cell growth, transfection, incubation/production, and recovery of the products. Inertial microfluidic cell manipulation processes allow many novel controls over host cells in bioreactors, including selective separation of transfected from untransfected host cells. Finally, a continuous bioprocessing system will be optimized to maximize the quantity and quality of the finished viral vector products. For this, we will employ novel microfluidic electrokinetic separation of empty from filled viral capsid, which can provide near real-time determination of the quality of the product. In contrast to fed-batch or batch cultures, continuous monitoring of vector quality and quantity enables rapid adjustment of the culture protocols to restore/maintain optimal conditions for vector production over an extended period. This project is expected to have a broader impact on the bioproduction of viral vectors, but especially on rare and ultra-rare diseases where low disease prevalence requires relatively small production volumes. Such diseases do not justify the biopharmaceutical industry’s large investment needed for vector manufacturing.

项目摘要摘要 高细胞密度无膜灌流培养连续生物生产病毒载体 在这个项目中，我们建议通过无膜灌流来强化病毒载体的生产过程。 生物反应器系统，这是我们在麻省理工学院的团队开发的。基于惯性微流体学和完全 在CHO细胞培养中得到验证，该系统比现有的灌流生物反应器具有许多关键优势， 包括高产品回收率和避免膜污染、连续清除死亡细胞、细胞碎片、 和蜂窝聚合，以及易于维护和自动化。因此，我们的生物反应器可以保持较高的细胞 即使细胞活力和培养条件发生显著变化，密度也很可靠，非常适合 控制病毒载体的生产过程。与基于HEK293和Sf9的生产专家合作 系统，我们将实施长期、高细胞密度的宿主细胞培养，并探索和验证几个 创新的载体生产平台，可更准确地控制新鲜细胞的生长、转基因、 产品的孵化/生产和回收。惯性微流控细胞操作过程允许 在生物反应器中对宿主细胞的许多新的控制，包括选择性地分离从 未转染的宿主细胞。最后，一个连续的生物处理系统将被优化，以最大化产量 以及成品病毒载体产品的质量。为此，我们将采用新型微流体动电学。 分离空的和填充的病毒衣壳，这可以提供近乎实时的质量测定 产品。与补料批次或批次培养不同，对病媒质量和数量的持续监测能够 快速调整培养方案，以恢复/保持最优的病媒生物生产条件 延长期限。该项目预计将对病毒载体的生物生产产生更广泛的影响，但 特别是罕见和超罕见疾病，这些疾病患病率较低，需要相对较少的生产 音量。这样的疾病并不能证明生物制药行业对病媒的大量投资是合理的。 制造业。