School of Chemistry, Manchester Institute of Biotechnology, University of Manchester, Manchester, M1 7DN, UK.
Analyst. 2013 Nov 21;138(22):6977-85. doi: 10.1039/c3an01341c.
UV resonance Raman (UVRR) spectroscopy combined with chemometric techniques was investigated as a physiochemical tool for monitoring secreted recombinant antibody production in cultures of Chinese hamster ovary (CHO) cells. Due to the enhanced selectivity of the UVRR, spectral variations arising from protein, small molecule substrates, and nucleic acid medium components could be measured simultaneously and we have successfully determined antibody titre. Medium samples were taken during culture of three CHO cell lines: two antibody-producing cell lines and a non-producing cell line, and analysed by UVRR spectroscopy using an excitation laser of 244 nm. Principal component analysis (PCA) was applied to the spectral sets and showed a linear trend over time for the antibody-producing cell lines that was not observed in the non-producing cell line. Partial least squares regression (PLSR) was used to predict antibody titres, glucose utilization and lactate accumulation, and compared very favourably with gold standard data acquired with the much slower techniques of ELISA and liquid chromatography. Further analysis of the UVRR spectral sets using two-dimensional correlation moving windows also revealed that spectral variations due to protein and nucleic acid concentrations in the medium during cell culture varied between each of the three cell lines investigated.
紫外共振拉曼（UVRR）光谱结合化学计量学技术被研究作为一种物理化学工具，用于监测中国仓鼠卵巢（CHO）细胞培养中分泌的重组抗体的生产。由于 UVRR 的增强选择性，可以同时测量蛋白质、小分子底物和核酸培养基成分引起的光谱变化，并且我们已经成功地确定了抗体效价。在三种 CHO 细胞系的培养过程中采集培养基样品：两种产生抗体的细胞系和一种不产生细胞系，并使用激发激光为 244nm 的 UVRR 光谱进行分析。主成分分析（PCA）应用于光谱集，并显示出抗体产生细胞系随时间的线性趋势，而在非产生细胞系中没有观察到这种趋势。偏最小二乘回归（PLSR）用于预测抗体效价、葡萄糖利用率和乳酸积累，与使用 ELISA 和液相色谱等较慢技术获得的金标准数据相比非常有利。使用二维相关移动窗口对 UVRR 光谱集进行的进一步分析还表明，在细胞培养过程中，培养基中蛋白质和核酸浓度引起的光谱变化在三个研究的细胞系之间有所不同。
