Multiple Reaction Monitoring Enables Precise Quantification of 97 Proteins in Dried Blood Spots

Simple item page
dc.contributor.authorChambers, Andrew G.
dc.contributor.authorPercy, Andrew J.
dc.contributor.authorYang, Juncong
dc.contributor.authorBorchers, Christoph H.
dc.date.accessioned2021-01-22T00:09:43Z
dc.date.available2021-01-22T00:09:43Z
dc.date.copyright2015en_US
dc.date.issued2015
dc.description.abstractThe dried blood spot (DBS) methodology provides a minimally invasive approach to sample collection and enables room-temperature storage for most analytes. DBS samples have successfully been analyzed by liquid chromatography multiple reaction monitoring mass spectrometry (LC/MRM-MS) to quantify a large range of small molecule biomarkers and drugs; however, this strategy has only recently been explored for MS-based proteomics applications. Here we report the development of a highly multiplexed MRM assay to quantify endogenous proteins in human DBS samples. This assay uses matching stable isotope-labeled standard peptides for precise, relative quantification, and standard curves to characterize the analytical performance. A total of 169 peptides, corresponding to 97 proteins, were quantified in the final assay with an average linear dynamic range of 207-fold and an average R2 value of 0.987. The total range of this assay spanned almost 5 orders of magnitude from serum albumin (P02768) at 18.0 mg/ml down to cholinesterase (P06276) at 190 ng/ml. The average intra-assay and inter-assay precision for 6 biological samples ranged from 6.1–7.5% CV and 9.5–11.0% CV, respectively. The majority of peptide targets were stable after 154 days at storage temperatures from −20 °C to 37 °C. Furthermore, protein concentration ratios between matching DBS and whole blood samples were largely constant (<20% CV) across six biological samples. This assay represents the highest multiplexing yet achieved for targeted protein quantification in DBS samples and is suitable for biomedical research applications.en_US
dc.description.reviewstatusRevieweden_US
dc.description.scholarlevelFacultyen_US
dc.identifier.citationChambers, A. G., Percy, A. J., Yang, J., & Borchers, C. H. (2015). Multiple Reaction Monitoring Enables Precise Quantification of 97 Proteins in Dried Blood Spots. Molecular & Cellular Proteomics, 14(11), 3094-3104. https://doi.org/10.1074/mcp.O115.049957.en_US
dc.identifier.urihttps://doi.org/10.1074/mcp.O115.049957
dc.identifier.urihttp://hdl.handle.net/1828/12580
dc.language.isoenen_US
dc.publisherMolecular & Cellular Proteomicsen_US
dc.subjectUVic Genome BC Proteomics Centre
dc.subject.departmentDepartment of Biochemistry and Microbiology
dc.titleMultiple Reaction Monitoring Enables Precise Quantification of 97 Proteins in Dried Blood Spotsen_US
dc.typeArticleen_US

Files

Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
Chambers_AndrewG_MolCellProteomics_2015.pdf
Size:
973.36 KB
Format:
Adobe Portable Document Format
Description:
Download
License bundle
Now showing 1 - 1 of 1
No Thumbnail Available
Name:
license.txt
Size:
2 KB
Format:
Item-specific license agreed upon to submission
Description:
Download

Collections

Faculty and Staff Publications
 
University of Victoria Libraries

We acknowledge and respect the Lək̓ʷəŋən (Songhees and Esquimalt) Peoples on whose territory the university stands, and the Lək̓ʷəŋən and W̱SÁNEĆ Peoples whose historical relationships with the land continue to this day.