National University of Ireland, Galway, Galway, Ireland.
Biotechnol Bioeng. 2010 Oct 1;107(2):290-301. doi: 10.1002/bit.22813.
The use of Raman spectroscopy coupled with chemometrics for the rapid identification, characterization, and quality assessment of complex cell culture media components used for industrial mammalian cell culture was investigated. Raman spectroscopy offers significant advantages for the analysis of complex, aqueous-based materials used in biotechnology because there is no need for sample preparation and water is a weak Raman scatterer. We demonstrate the efficacy of the method for the routine analysis of dilute aqueous solution of five different chemically defined (CD) commercial media components used in a Chinese Hamster Ovary (CHO) cell manufacturing process for recombinant proteins.The chemometric processing of the Raman spectral data is the key factor in developing robust methods. Here, we discuss the optimum methods for eliminating baseline drift, background fluctuations, and other instrumentation artifacts to generate reproducible spectral data. Principal component analysis (PCA) and soft independent modeling of class analogy (SIMCA) were then employed in the development of a robust routine for both identification and quality evaluation of the five different media components. These methods have the potential to be extremely useful in an industrial context for "in-house" sample handling, tracking, and quality control.
研究了拉曼光谱结合化学计量学在快速鉴定、表征和评估用于工业哺乳动物细胞培养的复杂细胞培养介质成分方面的应用。拉曼光谱在分析生物技术中使用的复杂、基于水的材料方面具有显著优势，因为不需要样品制备，并且水是弱拉曼散射体。我们证明了该方法在常规分析稀水溶液中的五种不同的化学定义（CD）商业介质成分方面的功效，这些成分用于中国仓鼠卵巢（CHO）细胞生产重组蛋白的过程中。化学计量学处理拉曼光谱数据是开发稳健方法的关键因素。在这里，我们讨论了消除基线漂移、背景波动和其他仪器artifact 的最佳方法，以生成可重复的光谱数据。然后，主成分分析（PCA）和软独立建模分类类比（SIMCA）被用于开发一种稳健的方法，用于鉴定和评估五种不同的培养基成分的质量。这些方法在工业环境中对于“内部”样品处理、跟踪和质量控制具有极大的潜力。
