title: Proteomic analysis of plasma derived from platelet buffy coats during storage at room temperature. An application of ProteoMiner™ technology.
abstract: The present study was aimed at revealing new insights into the analysis of storage-related processes occurring in the supernatants of platelet concentrates (PCs) derived from pooled buffy coats suspended in whole plasma. To reduce the dynamic range of plasma protein concentrations and access low-abundance proteins, we made use of a solid-phase combinatorial peptide ligand library, known under the trade name of ProteoMiner™. Afterwards, two-dimensional electrophoresis (2-DE) was coupled with mass spectrometry (MS) to reveal changes in proteomic profiles. Several storage-induced protein alterations were identified including changes to major plasma proteins. In particular, a precursor of the secretory form of clusterin was shown to accumulate during storage of PC supernatants, together with platelet-derived tropomyosin, suggesting a progressive loss of platelet integrity. Platelet-released proteins following activation have also been detected (alpha-1-B-glycoprotein, kininogen-1, and serpin proteinase inhibitor 8). Moreover, specific protein fragments (vitronectin, plakoglobin, hornerin, and apolipoprotein A-IV) were found to be modulated upon storage, possibly indicating a time-dependent buffy-coat PC deterioration. Globally, our findings provided the disclosure of unique proteins in PC supernatants with respect to previous studies conducted in similar experimental conditions, suggesting ProteoMiner enrichment technology to be a possible complementary tool in the identification of diagnostically relevant proteins as age/quality biomarkers of therapeutic products.
authors.affiliation: Department of Environmental Sciences, University of Tuscia, Viterbo, Italy.
authors.forename: Maria Giulia
authors.initials: MG
authors.lastname: Egidi
authors.forename: Sara
authors.initials: S
authors.lastname: Rinalducci
authors.forename: Cristina
authors.initials: C
authors.lastname: Marrocco
authors.forename: Stefania
authors.initials: S
authors.lastname: Vaglio
authors.forename: Lello
authors.initials: L
authors.lastname: Zolla
mesh_terms: D000595:Amino Acid Sequence; D058621:Blood Buffy Coat / Q000378:metabolism*; D001792:Blood Platelets / Q000378:metabolism*; D001793:Blood Preservation; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D040901:Proteomics; D016415:Sequence Alignment; D013696:Temperature; D019096:Vitronectin / Q000737:chemistry / Q000378:metabolism
chemical_list: D019096:Vitronectin