Metabolomics - a useful tool for prediction of protein production and processing?

Abstract

Although CHO cells are widely used as hosts for recombinant protein production, still only little is known regarding the interaction of metabolic alterations and protein production and processing. Therefore, a more sophisticated look into the cellular metabolism might lead to a more efficient use of the production medium resulting in high quantities of the protein with the desired product quality. For intracellular metabolite quantification the sample preparation including quenching of cells is a very critical step strongly affecting subsequent results. A simple and straightforward protocol, not needing special equipment, was investigated focusing on the efficiency of stopping metabolic activities and the potential metabolic leakage due to losses in membrane integrity. In a next attempt, the comparison between the producer cell line and the corresponding mock cell line using targeted mass spectrometry approaches for intra- and extracellular metabolite quantification was performed. The influence of the increased protein production capacity and the need of a complex glycosylation machinery on metabolite profiles was assessed. In order to find a link between the determined output parameters, detailed data evaluation including multivariate data analysis tools was implemented.