The stability of the circulating human proteome to variations in sample collection and handling procedures measured with an aptamer-based proteomics array.

Blood-based protein biomarkers hold great promise to advance medicine with applications that detect and diagnose diseases and aid in their treatment. We are developing such applications with our proteomics technology that combines high-content with low limits of detection. Biomarker discovery relies heavily on archived blood sample collections. Blood is dynamic and changes with different sampling procedures potentially confounding biomarker studies. In order to better understand the effects of sampling procedures on the circulating proteome, we studied three sample collection variables commonly encountered in archived sample sets. These variables included (1) three different sample tube types, PPT plasma, SST serum, and Red Top serum, (2) the time from venipuncture to centrifugation, and (3) the time from centrifugation to freezing. We profiled 498 proteins for each of 240 samples and compared the results by ANOVA. The results found no significant variation in the measurements for most proteins (approximately 99%) when the two sample processing times tested were 2h or less, regardless of sample tube type. Even at the longest timepoints, 20 h, approximately 82% of the proteins, on average for the three collection tube types, showed no significant change. These results are encouraging for proteomic biomarker discovery.

[1]  L. Gold,et al.  Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase. , 1990, Science.

[2]  G. Anderson,et al.  Effects of Blood Collection Conditions on Ovarian Cancer Serum Markers , 2007, PloS one.

[3]  L. Gold,et al.  Oligonucleotides as Research, Diagnostic, and Therapeutic Agents(*) , 1995, The Journal of Biological Chemistry.

[4]  Sen-Yung Hsieh,et al.  Systematical evaluation of the effects of sample collection procedures on low‐molecular‐weight serum/plasma proteome profiling , 2006, Proteomics.

[5]  N Leigh Anderson,et al.  High-abundance polypeptides of the human plasma proteome comprising the top 4 logs of polypeptide abundance. , 2008, Clinical chemistry.

[6]  Amanda Paulovich,et al.  Cancer biomarkers: a systems approach , 2006, Nature Biotechnology.

[7]  J. Neher A problem of multiple comparisons , 2011 .

[8]  J. Szostak,et al.  In vitro selection of RNA molecules that bind specific ligands , 1990, Nature.

[9]  Jaehoon Yu,et al.  An RNA aptamer that recognizes a specific conformation of the protein calsenilin. , 2007, Bioorganic & medicinal chemistry.

[10]  William E Grizzle,et al.  Standard operating procedures for serum and plasma collection: early detection research network consensus statement standard operating procedure integration working group. , 2009, Journal of proteome research.

[11]  S. Doberstein,et al.  HTS technologies in biopharmaceutical discovery. , 2006, Drug discovery today.

[12]  S. Jayasena Aptamers: an emerging class of molecules that rival antibodies in diagnostics. , 1999, Clinical chemistry.

[13]  Graham B. I. Scott,et al.  HUPO Plasma Proteome Project specimen collection and handling: Towards the standardization of parameters for plasma proteome samples , 2005, Proteomics.

[14]  John W. Tukey,et al.  Multiple comparisons: 1948-1983 , 1994 .

[15]  Sheela M. Waugh,et al.  2′-Fluoropyrimidine RNA-based Aptamers to the 165-Amino Acid Form of Vascular Endothelial Growth Factor (VEGF165) , 1998, The Journal of Biological Chemistry.

[16]  P. Maguire,et al.  Insights into the platelet releasate. , 2007, Current pharmaceutical design.

[17]  J. Schellens,et al.  Influence of variations in sample handling on SELDI‐TOF MS serum protein profiles for colorectal cancer , 2008, Proteomics. Clinical applications.

[18]  H. Verheul,et al.  Proteomics of the TRAP-induced platelet releasate. , 2009, Journal of proteomics.

[19]  B. Nilsson,et al.  Complement and coagulation: strangers or partners in crime? , 2007, Trends in immunology.

[20]  Zhiyuan Luo,et al.  Preanalytic influence of sample handling on SELDI-TOF serum protein profiles. , 2007, Clinical chemistry.

[21]  P. Siljander,et al.  Platelet collagen receptors and coagulation. A characteristic platelet response as possible target for antithrombotic treatment. , 2005, Trends in cardiovascular medicine.

[22]  N. Heegaard,et al.  Unravelling in vitro variables of major importance for the outcome of mass spectrometry-based serum proteomics. , 2007, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[23]  L. Williams,et al.  Discovery of a Cytokine and Its Receptor by Functional Screening of the Extracellular Proteome , 2008, Science.

[24]  L. Gold,et al.  The use of aptamers in large arrays for molecular diagnostics. , 1999, Molecular diagnosis : a journal devoted to the understanding of human disease through the clinical application of molecular biology.

[25]  Alex Stewart,et al.  Automation of the SomaLogic Proteomics Assay: A Platform for Biomarker Discovery , 2009 .

[26]  Akhilesh Pandey,et al.  Plasma Proteome Database as a resource for proteomics research , 2005, Proteomics.

[27]  R. Becker,et al.  Factor IXa inhibitors as novel anticoagulants. , 2007, Arteriosclerosis, thrombosis, and vascular biology.

[28]  Larry Gold,et al.  Proteomics and diagnostics: Let's Get Specific, again. , 2008, Current opinion in chemical biology.

[29]  Eugene A. Kapp,et al.  Overview of the HUPO Plasma Proteome Project: Results from the pilot phase with 35 collaborating laboratories and multiple analytical groups, generating a core dataset of 3020 proteins and a publicly‐available database , 2005, Proteomics.