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A Novel Approach for Using Dielectric Spectroscopy to Predict Viable Cell Volume (VCV) in Early Process Development

机译：在早期工艺开发中使用介电谱预测活细胞体积（VCV）的新方法

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Online monitoring of viable cell volume (VCV) is essential to the development, monitoring, and control of bioprocesses. The commercial availability of steam-sterilizable dielectric-spectroscopy probes has enabled successful adoption of this technology as a key noninvasive method to measure VCV for cell-culture processes. Technological challenges still exist, however. For some cell lines, the technique's accuracy in predicting the VCV from probe-permittivity measurements declines as the viability of the cell culture decreases. To investigate the cause of this decrease in accuracy, divergences in predicted vs. actual VCV measurements were directly related to the shape of dielectric frequency scans collected during a cell culture. The changes in the shape of the beta dispersion, which are associated with changes in cell state, are quantified by applying a novel "area ratio" (AR) metric to frequency-scanning data from the dielectric-spectroscopy probes. The AR metric is then used to relate the shape of the beta dispersion to single-frequency permittivity measurements to accurately predict the offline VCV throughout an entire fed-batch run, regardless of cell state. This work demonstrates the possible feasibility of quantifying the shape of the beta dispersion, determined from frequency-scanning data, for enhanced measurement of VCV in mammalian cell cultures by applying a novel shape-characterization technique. In addition, this work demonstrates the utility of using changes in the shape of the beta dispersion to quantify cell health.
机译：在线监测活细胞体积（VCV）对于开发，监视和控制生物过程至关重要。蒸汽灭菌介电谱探针的商业可得性已使该技术成功地用作测量细胞培养过程中VCV的关键非侵入性方法。但是，技术挑战仍然存在。对于某些细胞系，随着细胞培养物活力的降低，从探针介电常数测量值预测VCV的技术准确性也会下降。为了研究准确性降低的原因，预测VCV测量值与实际VCV测量值的差异与细胞培养过程中收集的介电频率扫描的形状直接相关。通过将新颖的“面积比”（AR）度量应用于介电谱探针的频率扫描数据，可以量化与细胞状态变化相关的β分散体形状变化。然后使用AR度量将β分散体的形状与单频介电常数测量值相关联，以准确预测整个补料分批运行中的离线VCV，而与细胞状态无关。这项工作证明了通过应用新型的形状表征技术，量化从频率扫描数据确定的β分散体形状的可行性，以增强哺乳动物细胞培养物中VCV的测量。此外，这项工作演示了使用β分散体形状的变化来量化细胞健康的实用性。

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