title: Identification of low abundant secreted proteins and peptides from primary culture supernatants of human T-cells.
abstract: Live cells continually communicate with their surroundings by the secretion of biomolecules, among which proteins and/or peptides are an important class. As such, these protein/peptide signals which end up in the extracellular medium, reflect the state of a cell in a certain condition, and as by definition are potential biomarkers indicative for specific physiological/pathological processes. We here report on a mass spectrometry based method for the detection and analysis of peptides and proteins secreted in a highly complex background, such as cell culture supernatant. Our method, which combines chromatography, high duty cycle tandem mass spectrometry and bio-informatics, enables the detection of interleukin-2 (IL-2), a cytokine secreted by activated T-cells, present in cell supernatant while representing only 0.006‰ of the total protein content. Moreover, the method allows the mass spectrometric analysis of signaling proteins in a non-targeted way and without any prior immunodepletion of the highest abundant cell culture medium proteins. In this study this is exemplified by the detection of yet two other secretory peptides, i.e., the granulins A and B, in the primary culture supernatant of non-activated T-cells.
authors.affiliation: Dept. of Biotechnology, Analytical Biotechnology Section, Delft University of Technology, Julianalaan 67, 2628BC, The Netherlands. i.m.o.finoulst@tudelft.nl
authors.forename: Inez
authors.initials: I
authors.lastname: Finoulst
authors.forename: Paul
authors.initials: P
authors.lastname: Vink
authors.forename: Eric
authors.initials: E
authors.lastname: Rovers
authors.forename: Mervin
authors.initials: M
authors.lastname: Pieterse
authors.forename: Martijn
authors.initials: M
authors.lastname: Pinkse
authors.forename: Ebo
authors.initials: E
authors.lastname: Bos
authors.forename: Peter
authors.initials: P
authors.lastname: Verhaert
mesh_terms: D017077:Culture Media, Conditioned / Q000494:pharmacology; D016207:Cytokines / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D010455:Peptides / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D011506:Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D013601:T-Lymphocytes / Q000166:cytology / Q000378:metabolism*; D053719:Tandem Mass Spectrometry
chemical_list: D017077:Culture Media, Conditioned; D016207:Cytokines; D010455:Peptides; D011506:Proteins