Nanoscale Biophotonics Laboratory, School of Chemistry, National University of Ireland, Galway, Galway, Ireland.
Appl Spectrosc. 2011 Nov;65(11):1240-9. doi: 10.1366/11-06383.
The application of fluorescence excitation-emission matrix (EEM) spectroscopy to the quantitative analysis of complex, aqueous solutions of cell culture media components was investigated. These components, yeastolate, phytone, recombinant human insulin, eRDF basal medium, and four different chemically defined (CD) media, are used for the formulation of basal and feed media employed in the production of recombinant proteins using a Chinese Hamster Ovary (CHO) cell based process. The comprehensive analysis (either identification or quality assessment) of these materials using chromatographic methods is time consuming and expensive and is not suitable for high-throughput quality control. The use of EEM in conjunction with multiway chemometric methods provided a rapid, nondestructive analytical method suitable for the screening of large numbers of samples. Here we used multiway robust principal component analysis (MROBPCA) in conjunction with n-way partial least squares discriminant analysis (NPLS-DA) to develop a robust routine for both the identification and quality evaluation of these important cell culture materials. These methods are applicable to a wide range of complex mixtures because they do not rely on any predetermined compositional or property information, thus making them potentially very useful for sample handling, tracking, and quality assessment in biopharmaceutical industries.
荧光激发-发射矩阵（EEM）光谱法在复杂的细胞培养介质成分水溶液的定量分析中的应用得到了研究。这些成分，酵母提取物、植物蛋白胨、重组人胰岛素、eRDF 基础培养基和四种不同的化学定义（CD）培养基，用于基于中国仓鼠卵巢（CHO）细胞的过程生产重组蛋白的基础培养基和补料培养基的配方。使用色谱方法对这些材料进行全面分析（无论是鉴定还是质量评估）既耗时又昂贵，并且不适合高通量质量控制。使用 EEM 结合多变量化学计量学方法提供了一种快速、非破坏性的分析方法，适用于大量样品的筛选。在这里，我们使用多向稳健主成分分析（MROBPCA）结合 n 向偏最小二乘判别分析（NPLS-DA），为这些重要的细胞培养材料的鉴定和质量评估开发了一种稳健的常规方法。这些方法适用于广泛的复杂混合物，因为它们不依赖于任何预定的组成或性质信息，因此在生物制药行业中，它们在样品处理、跟踪和质量评估方面具有很大的潜力。
