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首页> 外文期刊>Biotechnology Progress >Advanced Online Monitoring of Cell Culture Off-Gas Using Proton Transfer Reaction Mass Spectrometry
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Advanced Online Monitoring of Cell Culture Off-Gas Using Proton Transfer Reaction Mass Spectrometry

机译：使用质子转移反应质谱技术对细胞培养气体进行高级在线监测

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Mass spectrometry has been frequently applied to monitor the O2 and CO2 content in the off-gas of animal cell culture fermentations. In contrast to classical mass spectrometry the proton transfer reaction mass spectrometry (PTR-MS) provides additional information of volatile organic compounds by application of a soft ionization technology. Hence, the spectra show less fragments and can more accurately assigned to particular compounds. In order to discriminate between compounds of non-metabolic and metabolic origin cell free experiments and fed-batch cultivations with a recombinant CHO cell line were conducted. As a result, in total eight volatiles showing high relevance to individual cultivation or cultivation conditions could be identified. Among the detected compounds methanethiol, with a mass-to-charge ratio of 49, qualifies as a key candidate in process monitoring due to its strong connectivity to lactate formation. Moreover, the versatile and complex data sets acquired by PTR MS provide a valuable resource for statistical modeling to predict non direct measurable parameters. Hence, partial least square regression was applied to the complete spectra of volatiles measured and important cell culture parameters such as viable cell density estimated (R = 0.86). As a whole, the results of this study clearly show that PTR-MS provides a powerful tool to improve bioprocess-monitoring for mammalian cell culture. Thus, specific volatiles emitted by cells and measured online by the PTR-MS and complex variables gained through statistical modeling will contribute to a deeper process understanding in the future and open promising perspectives to bioprocess control.
机译：质谱法经常被用于监测动物细胞培养物发酵废气中的O2和CO2含量。与传统质谱相比，质子转移反应质谱（PTR-MS）通过应用软电离技术提供了挥发性有机化合物的其他信息。因此，光谱显示较少的碎片，可以更准确地分配给特定化合物。为了区分非代谢和代谢起源的化合物，进行了无细胞实验，并用重组CHO细胞系进行了分批补料培养。结果，总共可以鉴定出与个体栽培或栽培条件高度相关的八种挥发物。在检测到的化合物中，质荷比为49的甲硫醇由于其与乳酸形成的强连通性，因此可作为过程监控的关键候选物。此外，由PTR MS采集的通用和复杂数据集为统计建模提供了宝贵的资源，以预测不可直接测量的参数。因此，将偏最小二乘回归应用于所测挥发物的完整光谱以及重要的细胞培养参数，例如估计的活细胞密度（R = 0.86）。总体而言，这项研究的结果清楚地表明，PTR-MS提供了强大的工具来改善哺乳动物细胞培养的生物过程监测。因此，细胞排放的特定挥发物以及通过PTR-MS在线测量的挥发性物质以及通过统计建模获得的复杂变量将有助于将来对过程的更深入了解，并为生物过程控制打开广阔的前景。

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