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首页> 美国卫生研究院文献>Engineering in Life Sciences >A novel LED‐based 2D‐fluorescence spectroscopy system for in‐line monitoring of Chinese hamster ovary cell cultivations – Part I
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A novel LED‐based 2D‐fluorescence spectroscopy system for in‐line monitoring of Chinese hamster ovary cell cultivations – Part I

机译：一种新型的基于LED的二维荧光光谱系统用于在线监测中国仓鼠卵巢细胞的培养–第一部分

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A new two‐dimensional fluorescence sensor system was developed for in‐line monitoring of mammalian cell cultures. Fluorescence spectroscopy allows for the detection and quantification of naturally occurring intra‐ and extracellular fluorophores in the cell broth. The fluorescence signals correlate to the cells’ current redox state and other relevant process parameters. Cell culture pretests with twelve different excitation wavelengths showed that only three wavelengths account for a vast majority of spectral variation. Accordingly, the newly developed device utilizes three high‐power LEDs as excitation sources in combination with a back‐thinned CCD‐spectrometer for fluorescence detection. This setup was first tested in a lab design of experiments study with process relevant fluorophores proving its suitability for cell culture monitoring with LOD in the μg/L range. The sensor was then integrated into a CHO‐K1 cell culture process. The acquired fluorescence spectra of several batches were evaluated using multivariate methods. The resulting batch evolution models were challenged in deviating and “golden batch” validation runs. These first tests showed that the new sensor can trace the cells’ metabolic state in a fast and reliable manner. Cellular distress is quickly detected as a deviation from the “golden batch”.
机译：开发了一种新的二维荧光传感器系统，用于在线监测哺乳动物细胞培养物。荧光光谱法可以检测和定量细胞培养液中天然存在的细胞内和细胞外荧光团。荧光信号与细胞的当前氧化还原状态和其他相关过程参数相关。具有十二种不同激发波长的细胞培养物预测试表明，只有三种波长占据了光谱变化的绝大部分。因此，新开发的设备将三个大功率LED用作激发源，并结合了背面较薄的CCD光谱仪进行荧光检测。该设置首先在实验研究的实验室设计中进行了测试，并使用了与过程相关的荧光团，证明了其适用于μg/ L范围内LOD的细胞培养监测。然后将传感器集成到CHO-K1细胞培养过程中。使用多元方法评估了几批获得的荧光光谱。最终的批次演化模型在偏离和“黄金批次”验证运行中受到挑战。这些最初的测试表明，新传感器可以快速，可靠地追踪细胞的代谢状态。快速检测到细胞窘迫是与“黄金批次”的偏离。

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