Monitoring cell culture metabolites, including media components and cellular byproducts, during bio manufacturing is critical for gaining insights into cell growth, productivity and product quality. Historically, cell culture metabolite analysis was a complicated process requiring several orthogonal methods to cover the large number of metabolites with diverse properties over wide concentration ranges. These off-line analyses are time consuming and not suitable for real time bioreactor monitoring. In this study, we present a high-throughput LC-MS method with a 17-min cycle time that is capable of simultaneously monitoring 93 cell culture metabolites, including amino acids, nucleic acids, vitamins, sugars and others. This method has high precision and accuracy and has been successfully applied to the daily profiling of bioreactors and raw material qualification. Information obtained in these studies has been used to identify limiting amino acids during production, which guided adjustments to the feed strategy that prevented the potential misincorporation of amino acids. This type of metabolite profiling can be further utilized to build predictive process models for adaptive feedback control and pave the road for continuous manufacturing and real-time release testing.

在生物制造过程中监测细胞培养代谢物，包括培养基成分和细胞副产物，对于深入了解细胞生长，生产力和产品质量至关重要。从历史上看，细胞培养物代谢物分析是一个复杂的过程，需要几种正交方法才能涵盖在宽浓度范围内具有多种特性的大量代谢物。这些离线分析非常耗时，不适用于实时生物反应器监控。在这项研究中，我们提出了一种具有17分钟循环时间的高通量LC-MS方法，该方法能够同时监测93种细胞培养代谢产物，包括氨基酸，核酸，维生素，糖等。该方法具有很高的精度和准确性，已成功应用于生物反应器的日常分析和原料鉴定。这些研究中获得的信息已用于鉴定生产过程中的限制氨基酸，从而指导了对饲料策略的调整，以防止潜在的氨基酸错误掺入。这种类型的代谢物分析可以进一步用于建立预测过程模型以进行自适应反馈控制，并为连续生产和实时释放测试铺平道路。