Effective Quenching Processes for Physiologically Valid Metabolite Profiling of Suspension Cultured Mammalian Cells

Citations Over TimeTop 10% of 2008 papers

Abstract

Global metabolite analysis approaches, coupled with sophisticated data analysis and modeling procedures (metabolomics), permit a dynamic read-out of how cellular proteins interact with cellular and environmental conditions to determine cell status. This type of approach has profound potential for understanding, and subsequently manipulating, the regulation of cell function. As part of our study to define the regulatory events that may be used to maximize production of commercially valuable recombinant proteins from cultured mammalian cells, we have optimized the quenching process to allow retention of physiologically relevant intracellular metabolite profiles in samples from recombinant Chinese hamster ovary (CHO) cells. In a comparison of a series of candidate quenching procedures, we have shown that quenching in 60% methanol supplemented with 0.85% ammonium bicarbonate (AMBIC) at -40 degrees C generates a profile of metabolites that is representative of a physiological status based upon examination of key labile cellular metabolites. This represents a key feature for any metabolomic study with suspension cultured mammalian cells and provides confidence in the validity of subsequent data analysis and modeling procedures.

Related Papers

• → Establishment of a novel cell line, CHO-MK, derived from Chinese hamster ovary tissues for biologics manufacturing(2024)13 cited
• → Strategies for Efficient Transfection of CHO-Cells with Plasmid DNA(2011)6 cited
• → Metabolism of the neuroleptic agent zetidoline in the rat and the dog.(1984)4 cited
• Optimized Strategies to Hyperexpress Recombinant Protein in Chinese Hamster Ovary Cells.(2000)