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Title: Identification of low abundant secreted proteins and peptides from primary culture supernatants of human T-cells
Authors: Finoulst, Inez
Vink, Paul
Rovers, Eric
Pieterse, Mervin
Pinkse, Martijn
Bos, Ebo
Issue Date: 2011
Source: JOURNAL OF PROTEOMICS, 75(1), p. 23-33
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 parts per thousand 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.
Notes: [Finoulst, Inez; Pieterse, Mervin; Pinkse, Martijn; Verhaert, Peter] Delft Univ Technol, Analyt Biotechnol Sect, Dept Biotechnol, NL-2628 BC Delft, Netherlands. [Finoulst, Inez; Pieterse, Mervin; Pinkse, Martijn; Verhaert, Peter] Delft Univ Technol, Netherlands Prote Ctr, NL-2628 BC Delft, Netherlands. [Vink, Paul; Rovers, Eric; Bos, Ebo] MSD, Merck Res Labs, Dept Immune Therapeut, Oss, Netherlands. [Verhaert, Peter] Hasselt Univ, Biomed Res Inst, BIOMED, Diepenbeek, Belgium.
Keywords: Biochemical Research Methods; NanoLC; Tandem mass spectrometry; Secretome; Cytokines; Human lymphocytes;NanoLC; Tandem mass spectrometry; Secretome; Cytokines; Human lymphocytes
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ISSN: 1874-3919
e-ISSN: 1876-7737
DOI: 10.1016/j.jprot.2011.03.034
ISI #: 000298710400004
Category: A1
Type: Journal Contribution
Validations: ecoom 2013
Appears in Collections:Research publications

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