Neucode Labels for Multiplexed, Absolute Protein Quantification

We describe a new method to accomplish multiplexed, absolute protein quantification in a targeted fashion. The approach draws upon the recently developed neutron encoding (NeuCode) metabolic labeling strategy and parallel reaction monitoring (PRM). Since PRM scanning relies upon high-resolution tand...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2016-03, Vol.88 (6), p.3295-3303
Main Authors: Potts, Gregory K, Voigt, Emily A, Bailey, Derek J, Rose, Christopher M, Westphall, Michael S, Hebert, Alexander S, Yin, John, Coon, Joshua J
Format: Article
Language:English
Subjects:
Analytical chemistry
Cell Line, Tumor
Channels
Humans
Male
Mass spectra
Mass spectrometry
Mass spectroscopy
Monitoring
Multiplexing
Neutrons
Peptides
Proteins
Proteins - analysis
Scanning
Strategy
Tandem Mass Spectrometry
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Summary:We describe a new method to accomplish multiplexed, absolute protein quantification in a targeted fashion. The approach draws upon the recently developed neutron encoding (NeuCode) metabolic labeling strategy and parallel reaction monitoring (PRM). Since PRM scanning relies upon high-resolution tandem mass spectra for targeted protein quantification, incorporation of multiple NeuCode labeled peptides permits high levels of multiplexing that can be accessed from high-resolution tandem mass spectra. Here we demonstrate this approach in cultured cells by monitoring a viral infection and the corresponding viral protein production over many infection time points in a single experiment. In this context the NeuCode PRM combination affords up to 30 channels of quantitative information in a single MS experiment.
ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.5b04773