Real Time Cell Density Measurements During Production of Biologics and Other Biotech Products

 描述(由申请人提供)：这项第一阶段SBIR提案的目标是过程分析技术(PAT)中未得到满足的关键需求，即在生物制药的开发和制造过程中对细胞密度进行在线量化。提出了一种在基于酵母和哺乳动物细胞的生物过程中生产生物制品期间连续和实时操作的非破坏性的光学方法。这种方法利用了现有的创新生物过程监控器，该监控器可以在酵母(巴斯德毕赤酵母)和哺乳动物细胞(CHO)表达平台中自动测量生产过程中关键化学物质的浓度。使用近红外光谱方法，该测量系统在生物过程化学定量中表现出前所未有的稳定性，包括一年多的工业操作，不需要额外的校准。提出了一项研究计划，以确定使用该生物过程监测器对毕赤酵母和CHO细胞平台的细胞密度测量(细胞/毫升)进行量化的技术和商业可行性。这项建议的具体目标包括1.评估影响细胞密度定量的关键参数和可选的测量组合2.评估活蛋白表达生物过程中的校准稳健性。除了这项测量本身的高价值之外，定量细胞密度与关键营养和代谢物浓度的结合开辟了一系列令人印象深刻的基础和工程机会。例如，现在可以在复杂的生物过程操作期间实时评估和优化每个细胞的营养/代谢物利用的基本测量。这项技术有望在产品产量和产品一致性方面取得重大进展，从而降低这些复杂而有价值的药物的成本和上市时间。

 DESCRIPTION (provided by applicant): This Phase I SBIR proposal targets an unmet critical need in Process Analytical Technology (PAT), the on-line quantitation of cell density during the development and manufacture of biopharmaceuticals. A non-destructive, optical method is proposed that operates continuously and in real-time during production of biologics in yeast and mammalian cell-based bioprocesses. This approach leverages an existing innovative Bioprocess Monitor that automatically measures the concentration of key chemicals during production in yeast (Pichia pastoris) and mammalian cell (CHO) expression platforms. Using near-infrared spectroscopic methods, this measurement system has demonstrated unprecedented robustness in bioprocess chemical quantitation, including industrial operations over a year without need for additional calibration. A research plan is proposed to establish the technical and commercial feasibility of quantifying cell density measurements (cells/mL) using this Bioprocess Monitor for Pichia pastoris and CHO cell platforms. Specific Aims for this proposal include 1. Evaluation of critical parameters affecting cell density quantitation and alternative measurement combinations 2. Assessment of calibration robustness during live protein expression bioprocesses. In addition to the high value of this measurement by itself, the combination of quantitative cell density with key nutrient and metabolite concentrations opens an impressive range of fundamental and engineering opportunities. For example, fundamental measurements of nutrient/metabolite utilization per cell can now be evaluated and optimized in real-time during the complex bioprocess operation. Significant advances in product yield as well as product consistency are anticipated as a result of this technology, decreasing the cost and time to market for these complex and valuable medicines.