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 通过高细胞密度，无膜灌注培养物对病毒载体的连续生物生产 在这个项目中，我们建议通过无膜灌注来加强病毒载体的生产过程 生物反应器系统是由我们的团队在麻省理工学院开发的。基于惯性微流体和完全 该系统在Cho细胞种植中得到了验证，比现有的灌注生物反应器具有许多关键优势， 包括高产品恢复和避免膜结垢，连续去除死细胞，细胞碎片， 和细胞骨料，以及易于维护和自动化。结果，我们的生物反应器可以维持高细胞 即使细胞活力和培养条件发生显着变化，密度也可靠 控制病毒载体生产过程。与基于HEK293和SF9的专家合作 系统，我们将实施宿主细胞的长期高细胞密度培养，并探索和验证几个 创新的矢量生产平台可提供对新鲜细胞生长，翻译的更准确控制， 孵育/生产和产品的回收。惯性的微流体细胞操纵过程允许 对生物反应器中宿主细胞的许多新型控制，包括从中翻译的选择性分离 未转染的宿主细胞。最后，将优化连续的生物处理系统以最大化数量 和成品病毒矢量产品的质量。为此，我们将采用新型的微流体电动器 将空的病毒式衣壳分开，这可以几乎实时确定质量的质量 产品。与饲料批处理或批处理培养物相反，对向量质量和数量的连续监视可以实现 快速调整培养方案，以恢复/维持最佳条件，以使媒介生产在 长时间。预计该项目将对病毒载体的生物生产产生更大的影响，但 特别是在罕见和超稀有疾病的情况下，低疾病患病率需要相对较小的产生 卷。这样的疾病不能证明生物制药行业所需的大量投资 制造业。