RESEARCH ARTICLE Fast Filtration of Bacterial or Mammalian Suspension Cell Cultures for Optimal Metabolomics Results Natalie Bordag1, Vijay Janakiraman2, Jonny Nachtigall1, Sandra González Maldonado1, Bianca Bethan1, Jean-Philippe Laine3, Elie Fux1* 1 Metanomics GmbH, Berlin, Germany, 2 Biogen Idec Inc., Raleigh-Durham, North Carolina, United States of America, 3 Metanomics Health GmbH, Berlin, Germany a11111 *
[email protected] Abstract The metabolome offers real time detection of the adaptive, multi-parametric response of the OPEN ACCESS organisms to environmental changes, pathophysiological stimuli or genetic modifications and thus rationalizes the optimization of cell cultures in bioprocessing. In bioprocessing the Citation: Bordag N, Janakiraman V, Nachtigall J, González Maldonado S, Bethan B, Laine J-P, et al. measurement of physiological intracellular metabolite levels is imperative for successful (2016) Fast Filtration of Bacterial or Mammalian applications. However, a sampling method applicable to all cell types with little to no valida- Suspension Cell Cultures for Optimal Metabolomics tion effort which simultaneously offers high recovery rates, high metabolite coverage and Results. PLoS ONE 11(7): e0159389. doi:10.1371/ sufficient removal of extracellular contaminations is still missing. Here, quenching, centrifu- journal.pone.0159389 gation and fast filtration were compared and fast filtration in combination with a stabilizing Editor: Kyoung Heon Kim, Korea University, washing solution was identified as the most promising sampling method. Different influenc- REPUBLIC OF KOREA ing factors such as filter type, vacuum pressure, washing solutions were comprehensively Received: January 21, 2016 1 tested. The improved fast filtration method (MxP FastQuench) followed by routine lipid/ Accepted: July 3, 2016 polar extraction delivers a broad metabolite coverage and recovery reflecting well physio- Published: July 20, 2016 logical intracellular metabolite levels for different cell types, such as bacteria (Escherichia coli Copyright: © 2016 Bordag et al.