Rapid high-throughput characterisation, classification and selection of recombinant mammalian cell line phenotypes using intact cell MALDI-ToF mass spectrometry fingerprinting and PLS-DA modelling
Rapid high-throughput characterisation, classification and selection of recombinant mammalian cell line phenotypes using intact cell MALDI-ToF mass spectrometry fingerprinting and PLS-DA modelling
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DOI：
10.1016/j.jbiotec.2014.04.028
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发表时间：
2014-08-20
影响因子：
4.1
通讯作者：
Smales, C. Mark
中科院分区：
文献类型：
--
作者：
Povey, Jane F.;O'Malley, Christopher J.;Smales, C. Mark
尽管在过去几十年中在高产重组哺乳动物细胞系的产生方面取得了许多进展，但细胞系选择和开发通常由于无法使用来自小培养规模的早期细胞系构建的数据在较大规模（大体积生物反应器）下预测细胞系的表型特征（例如生长或重组蛋白生产率）而减慢。在这里，我们描述了一个完整的细胞MALDI-ToF质谱指纹的哺乳动物细胞在细胞系构建过程的早期方法的发展，由此产生的质谱数据被用来预测在更大的培养规模的哺乳动物细胞系的表型，使用偏最小二乘判别分析（PLS-DA）模型。使用MALDI-ToF质谱法，在细胞系开发的96深孔板阶段生成单个细胞系的质谱指纹库。在Lonza大规模（高达20，000 L）补料分批细胞培养工艺的10 L生物反应器模型中评价了这些细胞系的生长和生产率。使用96深孔板阶段的质谱信息和10 L生物反应器规模的表型信息，开发了PLS-DA模型，以基于96深孔板规模的MALDI-ToF指纹来预测10 L规模未知细胞系的生产力。该方法为cGMP生产规模生物反应器中细胞系性能的极早期预测提供了基础，并为从基于快速完整细胞质谱的测量中预测其他哺乳动物细胞表型的方法和模型提供了基础。(C)2014爱思唯尔有限公司版权所有。
Despite many advances in the generation of high producing recombinant mammalian cell lines over the last few decades, cell line selection and development is often slowed by the inability to predict a cell line's phenotypic characteristics (e.g. growth or recombinant protein productivity) at larger scale (large volume bioreactors) using data from early cell line construction at small culture scale. Here we describe the development of an intact cell MALDI-ToF mass spectrometry fingerprinting method for mammalian cells early in the cell line construction process whereby the resulting mass spectrometry data are used to predict the phenotype of mammalian cell lines at larger culture scale using a Partial Least Squares Discriminant Analysis (PLS-DA) model. Using MALDI-ToF mass spectrometry, a library of mass spectrometry fingerprints was generated for individual cell lines at the 96 deep well plate stage of cell line development. The growth and productivity of these cell lines were evaluated in a 10 L bioreactor model of Lonza's large-scale (up to 20,000 L) fed-batch cell culture processes. Using the mass spectrometry information at the 96 deep well plate stage and phenotype information at the 10 L bioreactor scale a PLS-DA model was developed to predict the productivity of unknown cell lines at the 10 L scale based upon their MALDI-ToF fingerprint at the 96 deep well plate scale. This approach provides the basis for the very early prediction of cell lines' performance in cGMP manufacturing-scale bioreactors and the foundation for methods and models for predicting other mammalian cell phenotypes from rapid, intact-cell mass spectrometry based measurements. (C) 2014 Elsevier B.V. All rights reserved.