Metabolic cytometry for in-line process analysis

项目摘要/摘要 基因疗法使用基因来预防和/或治疗获得性疾病和遗传性疾病。 近年来，对人类基因和相关疾病的深入研究提高了基因的能力 疗法是一种很有前途的未来疗法，可以显著延长数百万患者的预期寿命 患有不治之症的。这一前景加速了基因领域的指数增长。 心理治疗。然而，基因治疗的临床潜力仍然在很大程度上被释放，主要是由于有限的 临床和商业用基因传递产品的生物制造能力。基因制造 治疗性生物制剂涉及在高度复杂的哺乳动物细胞培养中获取病毒载体 而且很难控制。对病毒生产过程缺乏了解和控制导致了 制造挑战，包括生产率低、细胞线不稳定、杂质含量高以及 难以扩展，产品质量始终如一。因此，新的生物制造分析技术 是提高对生物过程动力学的理解和支持发展的关键 坚固而高效的生物制造工艺。 物理科学公司(PSI)与马萨诸塞大学洛厄尔分校(UML)合作提出了 开发一种新的生物制造过程分析技术工具，使在线、连续 治疗药物大规模生产用生物反应器中宿主细胞代谢的测定 病毒载体。一种新型的检测细胞内自体荧光的代谢细胞学传感器探针 代谢物将用于实时评估细胞水平的氧化还原代谢状态。创新的 空间和时间受限光谱方法被用于有效地区分荧光 来自非特异性光背景的细胞内代谢物信号，实现了高特异性 细胞生理学的测量。在第一阶段计划中，一种基于光纤的荧光原型 将制作和评估细胞测量探针系统，以展示实时的能力 用于病毒载体生产的生物反应器操作期间的代谢测量。技术成熟和 将进行在使用各种培养生物的附加生物反应器中的应用演示 在第二阶段计划期间。这项技术的成功开发将对 提高对大规模生产病毒载体的生物过程的理解和控制 基因治疗产品。

Project Summary/Abstract Gene therapy uses genes to prevent and/or treat acquired disorders and inherited genetic diseases. Recently, the intensive investigation of human genes and related diseases has improved the capability of gene therapy as a promising future therapy that can significantly increase life expectancy for millions of patients suffering from incurable diseases. This promise has accelerated the exponential growth in the field of gene therapy. However, the clinical potential of gene therapy remains largely unleashed, mainly due to the limited biomanufacturing capacity of gene delivery products for clinical and commercial use. Manufacturing of gene therapeutic biologics involves harvesting viral vectors in mammalian cell cultures which are highly complex and difficult to control. The lack of understanding and control of the viral production processes has led to manufacturing challenges including low productivity, instability of cell lines, high levels of impurities, and difficult scalability with consistent product quality. Therefore, new biomanufacturing analytical technologies are critically needed to improve the understanding of the bioprocess dynamics and to support development of robust and efficient biomanufacturing processes. Physical Sciences Inc. (PSI), in collaboration with the University of Massachusetts Lowell (UML) proposes to develop a novel biomanufacturing process analytical technology tool that enables in-line, continuous measurement of the metabolism of host cells in bioreactors for the large-scale manufacturing of therapeutic viral vectors. A novel metabolic cytometry sensor probe that measures the autofluorescence of intracellular metabolites will be used for real-time assessment of cellular-level redox metabolic state. An innovative spatially and temporally confined spectroscopy approach is used to efficiently differentiate fluorescence signals of intracellular metabolites from the nonspecific light background, achieving high specificity measurement of cellular physiology. During the Phase I program, a prototype fiber-based fluorescence cytometry probe system will be fabricated and evaluated, demonstrating the capability of the real-time metabolic measurement during bioreactor operations for viral vector production. Technology maturation and application demonstrations in additional bioreactors using a variety of culture organisms will be performed during the Phase II program. Successful development of this technology will contribute to significant improvement in the understanding and control of bioprocesses for large-scale manufacturing of viral vector products for gene therapy.