Multi-site assessment of the precision and reproducibility of multiple reaction monitoring-based measurements of proteins in plasma.
Verification of candidate biomarkers relies upon specific, quantitative assays optimized for selective detection of target proteins, and is increasingly viewed as a critical step in the discovery pipeline that bridges unbiased biomarker discovery to preclinical validation. Although individual laboratories have demonstrated that multiple reaction monitoring (MRM) coupled with isotope dilution mass spectrometry can quantify candidate protein biomarkers in plasma, reproducibility and transferability of these assays between laboratories have not been demonstrated. We describe a multilaboratory study to assess reproducibility, recovery, linear dynamic range and limits of detection and quantification of multiplexed, MRM-based assays, conducted by NCI-CPTAC. Using common materials and standardized protocols, we demonstrate that these assays can be highly reproducible within and across laboratories and instrument platforms, and are sensitive to low mug/ml protein concentrations in unfractionated plasma. We provide data and benchmarks against which individual laboratories can compare their performance and evaluate new technologies for biomarker verification in plasma.
Addona
Terri A
TA
Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
Abbatiello
Susan E
SE
Schilling
Birgit
B
Skates
Steven J
SJ
Mani
D R
DR
Bunk
David M
DM
Spiegelman
Clifford H
CH
Zimmerman
Lisa J
LJ
Ham
Amy-Joan L
AJ
Keshishian
Hasmik
H
Hall
Steven C
SC
Allen
Simon
S
Blackman
Ronald K
RK
Borchers
Christoph H
CH
Buck
Charles
C
Cardasis
Helene L
HL
Cusack
Michael P
MP
Dodder
Nathan G
NG
Gibson
Bradford W
BW
Held
Jason M
JM
Hiltke
Tara
T
Jackson
Angela
A
Johansen
Eric B
EB
Kinsinger
Christopher R
CR
Li
Jing
J
Mesri
Mehdi
M
Neubert
Thomas A
TA
Niles
Richard K
RK
Pulsipher
Trenton C
TC
Ransohoff
David
D
Rodriguez
Henry
H
Rudnick
Paul A
PA
Smith
Derek
D
Tabb
David L
DL
Tegeler
Tony J
TJ
Variyath
Asokan M
AM
Vega-Montoto
Lorenzo J
LJ
Wahlander
Asa
A
Waldemarson
Sofia
S
Wang
Mu
M
Whiteaker
Jeffrey R
JR
Zhao
Lei
L
Anderson
N Leigh
NL
Fisher
Susan J
SJ
Liebler
Daniel C
DC
Paulovich
Amanda G
AG
Regnier
Fred E
FE
Tempst
Paul
P
Carr
Steven A
SA
D015415:Biomarkers / Q000097:blood; D001774:Blood Chemical Analysis / Q000379:methods; D001798:Blood Proteins / Q000032:analysis*; D006801:Humans; D016014:Linear Models; D013058:Mass Spectrometry / Q000379:methods* / Q000592:standards; D020543:Proteome / Q000032:analysis; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013673:Technology Assessment, Biomedical
D015415:Biomarkers; D001798:Blood Proteins; D020543:Proteome