title: Simultaneous extraction of proteins and metabolites from cells in culture.
abstract: Proper sample preparation is an integral part of all omics approaches, and can drastically impact the results of a wide number of analyses. As metabolomics and proteomics research approaches often yield complementary information, it is desirable to have a sample preparation procedure which can yield information for both types of analyses from the same cell population. This protocol explains a method for the separation and isolation of metabolites and proteins from the same biological sample, in order for downstream use in metabolomics and proteomics analyses simultaneously. In this way, two different levels of biological regulation can be studied in a single sample, minimizing the variance that would result from multiple experiments. This protocol can be used with both adherent and suspension cell cultures, and the extraction of metabolites from cellular medium is also detailed, so that cellular uptake and secretion of metabolites can be quantified. Advantages of this technique includes:1.Inexpensive and quick to perform; this method does not require any kits.2.Can be used on any cells in culture, including cell lines and primary cells extracted from living organisms.3.A wide variety of different analysis techniques can be used, adding additional value to metabolomics data analyzed from a sample; this is of high value in experimental systems biology.
authors.affiliation: Luxembourg Centre for Systems Biomedicine, University of Luxembourg, L-4362 Esch-Belval, Luxembourg ; HICE - Helmholtz Virtual Institute of Complex Molecular Systems in Environmental Health - Aerosols and Health, Germany 1www.hice-vi.eu.
authors.forename: Sean C
authors.initials: SC
authors.lastname: Sapcariu
authors.affiliation: Mass Spectrometry Core Unit, Max-Delbrück Center for Molecular Medicine, 13125 Berlin, Germany ; HICE - Helmholtz Virtual Institute of Complex Molecular Systems in Environmental Health - Aerosols and Health, Germany 1www.hice-vi.eu.
authors.forename: Tamara
authors.initials: T
authors.lastname: Kanashova
authors.affiliation: Luxembourg Centre for Systems Biomedicine, University of Luxembourg, L-4362 Esch-Belval, Luxembourg.
authors.forename: Daniel
authors.initials: D
authors.lastname: Weindl
authors.affiliation: Luxembourg Centre for Systems Biomedicine, University of Luxembourg, L-4362 Esch-Belval, Luxembourg.
authors.forename: Jenny
authors.initials: J
authors.lastname: Ghelfi
authors.affiliation: Mass Spectrometry Core Unit, Max-Delbrück Center for Molecular Medicine, 13125 Berlin, Germany ; HICE - Helmholtz Virtual Institute of Complex Molecular Systems in Environmental Health - Aerosols and Health, Germany 1www.hice-vi.eu.
authors.forename: Gunnar
authors.initials: G
authors.lastname: Dittmar
authors.affiliation: Luxembourg Centre for Systems Biomedicine, University of Luxembourg, L-4362 Esch-Belval, Luxembourg ; HICE - Helmholtz Virtual Institute of Complex Molecular Systems in Environmental Health - Aerosols and Health, Germany 1www.hice-vi.eu.
authors.forename: Karsten
authors.initials: K
authors.lastname: Hiller
keywords: Biomolecule extraction; GC/MS; Isolation; LC/MS; Metabolomics; Proteomics; Sample preparation