Combining cell, metabolite, enzyme, and protein measurements with mathematical modeling generated a more holistic view on cellular processes involved in the adaptation of an adherent cell line to suspension growth.Īdaptation of mammalian cell lines to growth in suspension in a chemically defined medium has significant advantages for the design and optimization of manufacturing processes for biologicals. These regulatory mechanisms were linked to the proteins caveolin, cadherin-1, and pirin. Four regulatory mechanisms were identified as a response of the adaptation of adherent MDCK cells to growth in suspension. In addition, the cellular proteome was analyzed by liquid chromatography coupled to tandem mass spectrometry using a label-free protein quantification method to unravel altered cellular processes for the suspension and the adherent cell line. The experimental data set was used as the input for a segregated growth model that was already applied to describe the growth dynamics of the parental adherent cell line. Cell concentrations and cell volumes, extracellular metabolite concentrations, and intracellular enzyme activities were determined. Therefore, a Madin-Darby canine kidney (MDCK) suspension cell line, adapted earlier to growth in suspension, was cultivated in a 1-L bioreactor. In this study, for the first time, we combined several complex analytical approaches with the aim to track cellular changes on different levels and to unravel interconnections and correlations. So far, the complexity of alterations on the metabolism, enzyme, and proteome level required for adaptation is only poorly understood. Protein candidates to interfere glycolytic activity in MDCK cells.Īdaptations of animal cells to growth in suspension culture concern in particular viral vaccine production, where very specific aspects of virus-host cell interaction need to be taken into account to achieve high cell specific yields and overall process productivity.Concerted alteration of metabolic enzyme activity and protein expression.Less and more efficient glucose utilization for suspension cell growth.Combining cell, metabolite, enzyme, and protein measurements with mathematical modeling generated a more holistic view on cellular processes involved in the adaptation of an adherent cell line to suspension growth.
Adaptations of animal cells to growth in suspension culture concern in particular viral vaccine production, where very specific aspects of virus-host cell interaction need to be taken into account to achieve high cell specific yields and overall process productivity.
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