Exploring phlebotomy technique as a pre-analytical factor in proteomic analyses by mass spectrometry.
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.
a Department of Neurosciences, Stroke Rapid Assessment Unit (SRAU), Island Health, 1 Hospital Way, Victoria, BC V8Z 6R5, Canada.
Andrew M
AM
Penn
b Department of Research and Capacity Building, Island Health, 1952 Bay Street, Victoria, BC V8R 1J8, Canada.
Linghong
L
Lu
c University of Victoria, Genome British Columbia Proteomics Centre, Vancouver Island Technology Park, #3101 - 4464 Markham St., Victoria, BC V8Z 7X8, Canada.
Andrew G
AG
Chambers
d BC Centre for Disease Control, 655 West 12th Avenue, Vancouver, BC V5Z 4R4, Canada.
Robert F
RF
Balshaw
b Department of Research and Capacity Building, Island Health, 1952 Bay Street, Victoria, BC V8R 1J8, Canada.
Jaclyn L
JL
Morrison
b Department of Research and Capacity Building, Island Health, 1952 Bay Street, Victoria, BC V8R 1J8, Canada.
Kristine
K
Votova
e Department of Laboratory Medicine, Pathology and Medical Genetics, Island Health, 1952 Bay Street, Victoria, BC V8R 1J8, Canada.
Eileen
E
Wood
c University of Victoria, Genome British Columbia Proteomics Centre, Vancouver Island Technology Park, #3101 - 4464 Markham St., Victoria, BC V8Z 7X8, Canada.
Derek S
DS
Smith
b Department of Research and Capacity Building, Island Health, 1952 Bay Street, Victoria, BC V8R 1J8, Canada.
Maria
M
Lesperance
f Division of Hematology/Department of Medicine, Department of Neurology, University of Washington School of Medicine, 1959 N.E. Pacific Street, Seattle, WA 98195, USA.
Gregory J
GJ
del Zoppo
b Department of Research and Capacity Building, Island Health, 1952 Bay Street, Victoria, BC V8R 1J8, Canada.|c University of Victoria, Genome British Columbia Proteomics Centre, Vancouver Island Technology Park, #3101 - 4464 Markham St., Victoria, BC V8Z 7X8, Canada.|g Department of Biochemistry and Microbiology, University of Victoria, Petch Building Room 207, 3800 Finnerty Rd., Victoria, BC V8P 5C2, Canada.
Christoph H
CH
Borchers
D000328:Adult; D000368:Aged; D000704:Analysis of Variance; D015415:Biomarkers; D001772:Blood Cell Count; D001798:Blood Proteins; D004909:Erythrocyte Indices; D005260:Female; D006801:Humans; D008297:Male; D013058:Mass Spectrometry / Q000379:methods; D008875:Middle Aged; D018962:Phlebotomy / Q000379:methods; D025341:Principal Component Analysis; D040901:Proteomics / Q000379:methods
D015415:Biomarkers; D001798:Blood Proteins
biomarkers; biomarqueurs; mass spectrometry; multiple reaction monitoring; plasma; plasma sanguin; proteomics; protéomique; spectrométrie de masse; « multiple reaction monitoring » (MRM)