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Authors/year | Specimen | Target | Sample preparation | MS platform | Outcomes |
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Stahl-Zeng et al. 2007 [65] | Plasma
| N-glycoproteins | Selective isolation of N-glycosites. Stable isotope 13C- and/or 15N-labelled reference peptides. | LC ESI MS/MS Hybrid triple quadrupole linear ion trap | Detection ≤ 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 proteins | Top 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 proteins | Abundant 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 plasma | Immunodepletion (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) | NIL | ESI 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 protein | Immunodepletion of haptoglobin, IgG and HSA, then size exclusion chromatography | ESI 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. | PSA | Immunodepletion of albumin and mixed cation exchange peptide fractionation | ESI 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 | ESI LC-MS/MS 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) | NIL | ESI 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 33 | Immunoaffinity enrichment SISCAPA | ESI 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 21 | NIL | ESI 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 disease | NIL | ESI LC-MS/MS Agilent 6490 triple Quadrupole LC/MS | 117 from 135 peptides with attomolar LOQ for 81 peptides. |
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