Metabolic cytometry for in-line process analysis

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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.