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PeptiQuant™ Plus Kits

Multiplexed MRM-Based Protein Quantitation Using Two Different Stable Isotope-Labeled Peptide Isotopologues for Calibration

André LeBlanc,,Sarah A. Michaud,Andrew J. Percy,Darryl B. Hardie,Juncong Yang, Nicholas J. Sinclair,Jillaine I. Proudfoot,Adam Pistawka,Derek S. Smith, and Christoph H. Borchers*,,,§,,

† University of Victoria - Genome BC Proteomics Centre, Victoria, BC V8Z 7X8, Canada
‡ Proteomics Centre, Segal Cancer Centre, Lady Davis Institute, McGill University, Montreal, Quebec H3A 0G4, Canada
§ Department of Biochemistry and Microbiology, University of Victoria, Victoria, BC V8P 5C2, Canada
∥ Leibniz Institut für Analytische Wissenschaften - ISAS - e.V., Dortmund 44139, Germany
⊥ Gerald Bronfman Department of Oncology, Jewish General Hospital, Montreal, Quebec H3T 1E2, Canada

 


  • Multiplexed MRM-Based Protein Quantitation Using Two Different Stable Isotope-Labeled Peptide Isotopologues for Calibration
  • Inter-laboratory evaluation of instrument platforms and experimental workflows for quantitative accuracy and reproducibility assessment
  • Advances in multiplexed MRM-based protein biomarker quantitation toward clinical utility
  • A standardized kit for automated quantitative biomarker assessment of candidate protein biomarkers in human plasma
 
Researchers in academia and life science industries continue to adopt a targeted, bottom-up MS-based workflow for protein biomarker evaluation. Biomarker verification/validation requires absolute quantification of surrogate peptides in the sample matrix, a requirement that is best achieved using well-characterized standards. To ensure robust quantitative measurement, QC checks should be routinely performed.
 
CIL is pleased to offer a collection of PeptiQuant™ Assay Kits from MRM Proteomics Inc. for QC and biomarker assessment using bottom-up LC-MS/MS methodologies. The QC kits are designed to evaluate the performance of a LC-MS platform, either alone or in combination with a human or mouse plasma proteomic workflow. The biomarker assessment kits (BAKs) are intended to help researchers screen target panels of candidate protein disease biomarkers in human or mouse plasma samples.
 
Regarding the methodology, the original PeptiQuant kits utilize stable isotope-labeled standard (SIS) peptide mixtures, while the new PeptiQuant Plus kits employ mixtures of synthetic SIS and natural (NAT) peptides in the post-digestion spike of sample or surrogate matrices. Quantitation is achieved via reverse standard curves in the original kits and forward curves in the Plus kits (approaches described in PMID: 27341553). Comparatively, the new PeptiQuant Plus kits provide improved depth and precision of targeted protein quantitation. Further, these have been rigorously characterized according to the complete set of CPTAC (Clinical Proteomic Tumor Analysis Consortium) guidelines and are available for viewing on the CPTAC Assay Portal.
 

 

Resources

PeptiQuant™ Plus QC Flyer
PeptiQuant™ Plus BAK Flyer

PeptiQuant™ Plus BAK Panels

Stable Isotope-Labeled Peptide and Protein and Protein Reagents and Kits

 

For further information on these products, please visit MRM Proteomics, Inc. at  http://www.mrmproteomics.com

 

 

Frequently Asked Questions 

How many calibration curves can be prepared with a PeptiQuant™ Plus BAKs?

The SIS and NAT mixes enable 8-point curves that span a 1000-fold concentration range.  Each calibrant level is to be measured in singleton.
 

How many peptides/protein and MRM transitions do the PeptiQuant Plus kits monitor?

For each kit, one peptide/protein and three transitions per peptide.

Are the targets in the original PeptiQuant BAKs all present in the PeptiQuant Plus BAKs?

No. In the human plasma Plus BAK, 56 of the 76 proteins and 41 of the 101 peptides in the original kit are present. In contrast, 62 of the 81 proteins and 50 of the 101 peptides in the original kit are represented in the mouse plasma Plus BAK.

What types of disease associations have the proteins in the human plasma PeptiQuant Plus BAK been linked to?

The 125 proteins have been linked to innumerable diseases (e.g., 60 to cancer, 56 to cardiovascular disease, 39 to respiratory tract disease). Specific linkages (with references) can be provided upon request.

 

References

LeBlanc, A.; Michaud, S.A.; Percy, A.J.; et al. 2017. Multiplexed MRM-based protein quantitation using two different stable isotope-labeled peptide isotopologues for calibration. J Proteome Res, 16(7), 2527-2536. PMID: 28516774

Percy, A.J.; Michaud, S.A.; Jardim, A. 2016. Multiplexed MRM-based assays for the quantitation of proteins in mouse plasma and heart tissue. Proteomics, 17(7). PMID: 27688154

Hirtz,C.; Vialaret, J.; Nowak, N. 2016. Absolute quantification of 35 plasma biomarkers in human saliva using targeted MS. Bioanalysis, 8(1), 43-53. PMID: 26647646

Percy, A.J.; Mohammed, Y.; Yang, J. 2015. A standard kit for automated quantitative assessment of candidate protein biomakers in human plasma. Bioanalysis, 7(23), 2991-3004. PMID: 26626236

Mohammed, Y.; Percy, A.J.; Chambers, A.G. 2015. Qualis-SIS: automated standard curve generation and quality assessment for multiplexed targeted quantitative proteomic experiments with labeled standards. J Proteome Res, 14(2), 1137-1146. PMID: 25546269

Percy, A.J.; Simon, R.; Chambers, A.G.; Borchers, C.H. 2014. Enhanced sensitivity and mulitplexing with 2D LC/MRM-MS and labeled standards for deeper and more comprehensive protein quantitation. J Proteomics, 106, 113-124. PMID: 24769237

Percy, A.J.; Chambers, A.G.; Yang, J. 2013. Method and platform standardization in MRM-based quantitative plasma proteomics. J Proteomics, 95, 66-76. PMID: 23933160

Percy, A.J.; Chambers, A.G.; Smith, D.S. 2013. Standardized protocols for quality control of MRM-based plasma proteomic workflows. J Proteome Res, 12(1), 222-233. PMID: 23245390

Andrew Percy, PhD

Andrew Percy, PhD

Senior Applications Chemist – Mass Spectrometry

Dr. Andrew Percy is the Senior Applications Chemist for Mass Spectrometry. His responsibilities minimally involve overseeing product development, identifying new product market opportunities, assisting in the analysis of products for MS ‘omics applications, and providing technical support to customers.

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Kevin Millis, PhD

Kevin Millis, PhD

Senior Scientist, Application Development Manager

Kevin Millis, PhD, is the Senior Scientist and Market Development Manager for all NMR and Mass Spectrometry product lines. Kevin is responsible for Technical Services both internally and externally for all CIL customers as well as being responsible for the application and market development for the CIL products.

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