Penn, Andrew M., Lu, Linghong, Chambers, Andrew G., Balshaw, Robert F., Morrison, Jaclyn L., Votova, Kristine, Wood, Eileen, Smith, Derek S., Lesperance, Maria, del Zoppo, Gregory J., and Borchers, Christoph H.
Multiple reaction monitoring mass spectrometry (MRM-MS) is an emerging technology for blood biomarker verification and validation; however, the results may be influenced by pre-analytical factors. This exploratory study was designed to determine if differences in phlebotomy techniques would significantly affect the abundance of plasma proteins in an upcoming biomarker development study. Blood was drawn from 10 healthy participants using four techniques: (1) a 20-gauge IV with vacutainer, (2) a 21-gauge direct vacutainer, (3) an 18-gauge butterfly with vacutainer, and (4) an 18-gauge butterfly with syringe draw. The abundances of a panel of 122 proteins (117 proteins, plus 5 matrix metalloproteinase (MMP) proteins) were targeted by LC/MRM-MS. In addition, complete blood count (CBC) data were also compared across the four techniques. Phlebotomy technique significantly affected 2 of the 11 CBC parameters (red blood cell count, p = 0.010; hemoglobin concentration, p = 0.035) and only 12 of the targeted 117 proteins (p < 0.05). Of the five MMP proteins, only MMP7 was detectable and its concentration was not significantly affected by different techniques. Overall, most proteins in this exploratory study were not significantly influenced by phlebotomy technique; however, a larger study with additional patients will be required for confirmation. Key words: mass spectrometry, biomarkers, proteomics, plasma, multiple reaction monitoring. La spectrometrie de masse en mode MRM (<>; MRM-MS) est une technologie emergente pour la mesure et la validation de biomarqueurs sanguins, cependant ses resultats peuvent etre affectes par plusieurs facteurs pre-analytiques. Cette etude exploratoire a ete concue pour determiner si des differences dans les techniques de phlebotomie avaient un effet significatif sur l'abondance des proteines plasmatiques en vue d'un travail futur visant le developpement d'un biomarqueur. Du sang a ete preleve chez 10 participants en sante au moyen de quatre techniques : (1) IV 20 gauges sous vide (<>), (2) 21 gauges sous vide direct, (3) aiguille papillon 18 gauges sous vide, et (4) aiguille papillon a 18 gauges avec seringue. L'abondance de 122 proteines (117 proteines et 5 metalloproteinases matricielles (MMP)) a ete mesuree par LC/MRM-MS. En plus, les donnees issues de l'analyse de la formule sanguine complete (CBC) ont ete comparees pour les quatre techniques de prelevement. La technique de phlebotomie a affecte significativement deux des 11 parametres de la formule sanguine (nombre de globules rouges, p = 0,010; concentration de l'hemoglobine, p = 0,035) et seulement 12 des 117 proteines ciblees (p < 0,05). Des cinq MMR, seule MMP7 s'est averee detectable mais sa concentration n'etait pas affectee par la technique de prelevement sanguin. Globalement, la plupart des proteines examinees dans cette etude preliminaire n'etaient pas affectees par la technique de phlebotomie; cependant, une etude de plus grande ampleur comportant plus de patients sera necessaire pour en confirmer les resultats. [Traduit par la Redaction] Mots-cles: spectrometrie de masse, biomarqueurs, proteomique, plasma sanguin, <> (MRM)., Introduction Blood is a desirable sample for biomarker development because its collection is routine in most general, acute, and urgent care health settings. Multiple reaction monitoring mass spectrometry (MRM-MS) combined [...]