Direct quantitation of MHC-bound peptide epitopes by selected reaction monitoring

Chor Teck Tan, Nathan P Croft, Nadine L Dudek, Nicholas Andrew Williamson, Anthony W Purcell

Research output: Contribution to journalArticleResearchpeer-review

Abstract

We describe a cell-free approach that employs selected reaction monitoring (SRM) in tandem mass spectrometry to identify and quantitate T-cell epitopes. This approach utilises multiple epitope-specific SRM transitions to identify known T-cell epitopes and an absolute quantitation (AQUA) peptide strategy to afford AQUA. The advantage of a mass spectrometry-based approach over more traditional cell-based assays resides in the robustness and transferability of an SRM approach between laboratories and the ability of this strategy to detect multiple peptides simultaneously without the requirement of epitope-specific reagents such as T-cell lines. Thus, the SRM strategy for epitope quantitation will find application in studies of antigen density, the link between epitope abundance and immunogenicity, the dynamic range of epitope presentation and the abundance of T-cell epitopes in disease. ? 2011 WILEY-VCH Verlag GmbH Co. KGaA, Weinheim.
Original languageEnglish
Pages (from-to)2336 - 2340
Number of pages5
JournalProteomics
Volume11
Issue number11
DOIs
Publication statusPublished - 2011
Externally publishedYes

Cite this

Tan, Chor Teck ; Croft, Nathan P ; Dudek, Nadine L ; Williamson, Nicholas Andrew ; Purcell, Anthony W. / Direct quantitation of MHC-bound peptide epitopes by selected reaction monitoring. In: Proteomics. 2011 ; Vol. 11, No. 11. pp. 2336 - 2340.
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Direct quantitation of MHC-bound peptide epitopes by selected reaction monitoring. / Tan, Chor Teck; Croft, Nathan P; Dudek, Nadine L; Williamson, Nicholas Andrew; Purcell, Anthony W.

In: Proteomics, Vol. 11, No. 11, 2011, p. 2336 - 2340.

Research output: Contribution to journalArticleResearchpeer-review

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AU - Croft, Nathan P

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AB - We describe a cell-free approach that employs selected reaction monitoring (SRM) in tandem mass spectrometry to identify and quantitate T-cell epitopes. This approach utilises multiple epitope-specific SRM transitions to identify known T-cell epitopes and an absolute quantitation (AQUA) peptide strategy to afford AQUA. The advantage of a mass spectrometry-based approach over more traditional cell-based assays resides in the robustness and transferability of an SRM approach between laboratories and the ability of this strategy to detect multiple peptides simultaneously without the requirement of epitope-specific reagents such as T-cell lines. Thus, the SRM strategy for epitope quantitation will find application in studies of antigen density, the link between epitope abundance and immunogenicity, the dynamic range of epitope presentation and the abundance of T-cell epitopes in disease. ? 2011 WILEY-VCH Verlag GmbH Co. KGaA, Weinheim.

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