Use of stable isotope dimethyl labeling coupled to selected reaction monitoring to enhance throughput by multiplexing relative quantitation of targeted proteins

Thin Thin Aye, Low Teck Yew, Yngvild Bjørlykke, Harald Barsnes, Albert J R Heck, Frode S. Berven

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

In this manuscript, we present a proof-of-concept study for targeted relative protein quantitation workflow using chemical labeling in the form of dimethylation, coupled with selected reaction monitoring (dimethyl-SRM). We first demonstrate close to complete isotope incorporation for all peptides tested. The accuracy, reproducibility, and linear dynamic range of quantitation are further assessed based on known ratios of nonhuman standard proteins spiked into human cerebrospinal fluid (CSF) as a model complex matrix. Quantitation reproducibility below 20% (CV < 20%) was obtained for analyte concentrations present at a dynamic range of 4 orders of magnitude lower than that of the background proteins. An error of less than 15% was observed when measuring the abundance of 44 out of 45 major human plasma proteins. Dimethyl-SRM was further examined by comparing the relative quantitation of eight proteins in human CSF with the relative quantitation obtained using synthetic heavy peptides coupled to stable isotope dilution-SRM (SID-SRM). Comparison between the two methods reveals that the correlation between dimethyl-SRM and SID-SRM is within 0.3-33% variation, demonstrating the accuracy of relative quantitation using dimethyl-SRM. Dimethyl labeling coupled with SRM provides a fast, convenient, and cost-effective alternative for relative quantitation of a large number of candidate proteins/peptides.

Original languageEnglish
Pages (from-to)4999-5006
Number of pages8
JournalAnalytical Chemistry
Volume84
Issue number11
DOIs
Publication statusPublished - 5 Jun 2012
Externally publishedYes

Fingerprint

Multiplexing
Isotopes
Labeling
Throughput
Monitoring
Cerebrospinal fluid
Proteins
Peptides
Dilution
Blood Proteins
Costs

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Use of stable isotope dimethyl labeling coupled to selected reaction monitoring to enhance throughput by multiplexing relative quantitation of targeted proteins. / Aye, Thin Thin; Teck Yew, Low; Bjørlykke, Yngvild; Barsnes, Harald; Heck, Albert J R; Berven, Frode S.

In: Analytical Chemistry, Vol. 84, No. 11, 05.06.2012, p. 4999-5006.

Research output: Contribution to journalArticle

Aye, Thin Thin ; Teck Yew, Low ; Bjørlykke, Yngvild ; Barsnes, Harald ; Heck, Albert J R ; Berven, Frode S. / Use of stable isotope dimethyl labeling coupled to selected reaction monitoring to enhance throughput by multiplexing relative quantitation of targeted proteins. In: Analytical Chemistry. 2012 ; Vol. 84, No. 11. pp. 4999-5006.
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