TY - JOUR
T1 - PICquant
T2 - A quantitative platform to measure differential peptide abundance using dual-isotopic labeling with 12C6-and 13C6-phenyl isocyanate
AU - Lyons, Charles E.
AU - Victor, Ken G.
AU - Moshnikov, Sergey A.
AU - Bachmann, Lorin M.
AU - Baras, Alexander S.
AU - Dettmann, Kathleen M.
AU - Cross, Janet V.
AU - Templeton, Dennis J.
PY - 2011/2/1
Y1 - 2011/2/1
N2 - We have developed a complete system for the isotopic labeling, fractionation, and automated quantification of differentially expressed peptides that significantly facilitates candidate biomarker discovery. We describe a new stable mass tagging reagent pair, 12C6-and 13C6-phenyl isocyanate (PIC), that offers significant advantages over currently available tags. Peptides are labeled predominantly at their amino termini and exhibit elution profiles that are independent of label isotope. Importantly, PIC-labeled peptides have unique neutral-mass losses upon CID fragmentation that enable charge state and label isotope identification and, thereby, decouple the sequence identification from the quantification of candidate biomarkers. To exploit these properties, we have coupled peptide fractionation protocols with a Thermo LTQ-XL LC-MS2 data acquisition strategy and a suite of automated spectrum analysis software that identifies quantitative differences between labeled samples. This approach, dubbed the PICquant platform, is independent of protein sequence identification and excludes unlabeled peptides that otherwise confound biomarker discovery. Application of the PICquant platform to a set of complex clinical samples showed that the system allows rapid identification of peptides that are differentially expressed between control and patient groups.
AB - We have developed a complete system for the isotopic labeling, fractionation, and automated quantification of differentially expressed peptides that significantly facilitates candidate biomarker discovery. We describe a new stable mass tagging reagent pair, 12C6-and 13C6-phenyl isocyanate (PIC), that offers significant advantages over currently available tags. Peptides are labeled predominantly at their amino termini and exhibit elution profiles that are independent of label isotope. Importantly, PIC-labeled peptides have unique neutral-mass losses upon CID fragmentation that enable charge state and label isotope identification and, thereby, decouple the sequence identification from the quantification of candidate biomarkers. To exploit these properties, we have coupled peptide fractionation protocols with a Thermo LTQ-XL LC-MS2 data acquisition strategy and a suite of automated spectrum analysis software that identifies quantitative differences between labeled samples. This approach, dubbed the PICquant platform, is independent of protein sequence identification and excludes unlabeled peptides that otherwise confound biomarker discovery. Application of the PICquant platform to a set of complex clinical samples showed that the system allows rapid identification of peptides that are differentially expressed between control and patient groups.
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U2 - 10.1021/ac102461e
DO - 10.1021/ac102461e
M3 - Article
C2 - 21192683
AN - SCOPUS:79952143091
SN - 0003-2700
VL - 83
SP - 856
EP - 865
JO - Analytical Chemistry
JF - Analytical Chemistry
IS - 3
ER -