International Journal of Proteomics / 2013 / Article / Tab 3

Review Article

Current Status and Advances in Quantitative Proteomic Mass Spectrometry

Table 3

Summary of MRM quantitative analysis in blood for a variety disease types.

Authors/yearSpecimenTargetSample preparationMS platformOutcomes

Stahl-Zeng et al. 2007 [65]Plasma
N-glycoproteinsSelective isolation of N-glycosites. Stable isotope 13C- and/or 15N-labelled reference peptides.LC ESI MS/MS Hybrid triple quadrupole linear ion trapDetection ≤ ng/mL concentration range and accurate quantification over a linear range of ~ 105.
LOQ of 50 amol. LOD ≥ 10 amol. Protein concentration in plasma of 0.1 ng/mL.
Anderson and Hunter 2006 [66]Plasma
1 healthy donor
53 plasma proteinsTop six abundant proteins depleted. Stable isotope labeled internal standards.ESI LC-MS/MS
4000 Q Trap Hybrid triple quadrupole/linear ion trap
Quantitative data for 47 proteins in the µg/mL level over linear range of 104
Keshishian et al. 2007 [67]Plasma
1 healthy donor
6 low abundance plasma proteinsAbundant protein depletion and SCX chromatography. Stable isotope-labeled amino acids.ESI LC-MS/MS
4000 Q Trap Hybrid triple quadrupole/linear ion trap instrument
LOQ of 1–10 ng/mL range and linearity ≥ 102. LOD in high pg/mL.
McKay et al. 2007 [68]Plasma
4 colorectal cancer patients undergoing chemotherapy
18 liver-derived proteins in plasmaImmunodepletion (Albumin and IgG removed)ESI LC-MS/MS
4000 Q Trap Hybrid triple quadrupole/linear ion trap instrument
Increase in target plasma proteins during treatment. Similar trends found in MRM assays and 2-D DIGE
Kirsch et al. 2007 [69]Blood bank pooled serum
2 human growth hormones (IGFBP-3, IGF-1)NILESI LC-MS/MS
Triple quadrupole mass spectrometer
Detection ranges of 4–10 ng/µl for IGFBP-3 and 2–8 ng/µl for IGF-1.
Kuhn et al. 2004 [70]Serum.
Pools of healthy, non-erosive RA and erosive RA ( 5 individuals per pool).
C-reactive proteinImmunodepletion of haptoglobin, IgG and HSA, then size exclusion chromatographyESI LC-MS/MS
Triple quadrupole mass spectrometer
Correlation between erosive RA, RA and increased CRP over healthy patients.
Results verified using immunoassay.
Fortin et al. 2009 [71]Serum
Benign prostate hyperplasia and prostate cancer.
PSAImmunodepletion of albumin and mixed cation exchange peptide fractionationESI LC-MS/MS
API 2000 triple quadrupole or 4000 Q Trap hybrid triple quadrupole/linear ion trap
Absolute quant. of PSA to low ng/mL, with good correlation to clinical ELISA tests.
Huillet et al. 2012 [72]Serum
Clinical samples from 5 myocardial infarction patients
Clinically validated cardiovascular biomarkers (LDH-B, CKMB, myoglobin, troponin I)Immunodepletion of six highest abundant proteins and SDS-PAGE
Immunocapture prefractionation and SDS-PAGE
5500 Q Trap hybrid triple quadrupole/linear ion trap mass spectrometer
Absolute quant. using Protein Standard Absolute Quantification (PSAQ) and MRM.
Demonstrated good correlation with ELISA assay results.
Zhao et al. 2010 [73]Serum
10 hepatocellular carcinoma patients and 10 healthy donors
Candidate biomarkers of hepatocellular carcinoma (vitronectin and clusterin)NILESI LC-MS/MS
4000 Q Trap Hybrid triple quadrupole/linear ion trap instrument
Stable isotope dilution-MRM using 18O-labelling method demonstrated significant downregulated in HCC compared to healthy group. Results comparable to ELISA.
Kuhn et al. 2009 [74]Plasma
5 patients undergoing PMI and alcohol ablation treatment for HOCM
Troponin I, and Interleukin 33Immunoaffinity enrichment SISCAPAESI LC-MS/MS
Triple quadrupole instrument
Linearity from1.5 to 5000 µg/L and correlated with commercial immunoassay.
Demonstrated how SISCAPA-MRM can quantify changes to low µg/L levels.
Lopez et al. 2011 [75]Serum
from 24 trisomy 21 and 21 normal first trimester pregnancies
12 putative markers of Trisomy 21NILESI LC-MS/MS
LTQ Orbitrap XL mass spectrometer
Developed a workflow for Trisomy 21. Protein biomarkers targeted are high abundance proteins.
SRM LOQ of 1–5 femtomoles..
Domanski et al. 2012 [76]Plasma
90 patients with cardiovascular disease
67 putative markers of cardiovascular diseaseNILESI LC-MS/MS
Agilent 6490 triple Quadrupole LC/MS
117 from 135 peptides with attomolar LOQ for 81 peptides.