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Enhancing Data Reliability in TOMAHAQ for Large-Scale Protein Quantification
Fang Liu1, Kratika Singhal1, Rowan Matney1
1Vincent Coates Foundation Mass Spectrometry Laboratory, Stanford University, Stanford, CA, 94305, USA.
Proteomics
|February 8, 2020
Summary
The triggered by offset, multiplexed, accurate mass, high resolution, and absolute quantitation (TOMAHAQ) technique enhances proteomics assays. A new data correction strategy resolves interference from TMTzero reporter ions for improved quantification.
Area of Science:
- Proteomics
- Mass Spectrometry
- Quantitative Biology
Background:
- Targeted proteomics assays face limitations in analytical scale and instrument utilization.
- The TOMAHAQ technique was developed to improve both analytical scale and quantitative performance through peptide and sample multiplexing.
Purpose of the Study:
- To discuss critical technical requirements and data analysis for TOMAHAQ implementation.
- To address interference issues observed with TMTzero reporter ions in TOMAHAQ.
- To propose a data correction strategy for reliable high-throughput quantification.
Main Methods:
- Studied 185 target peptides across over 200 clinical plasma samples.
- Investigated interference from TMTzero reporter ions in TOMAHAQ assays.
- Developed a post-acquisition data correction strategy to deconvolute reporter ion superposition.
Main Results:
- Observed significant interference from TMTzero reporter ions, which was unexpected.
- Identified the source of interference as synthetic trigger peptides.
- Demonstrated the effectiveness of the proposed data correction strategy in recovering reliable data.
Conclusions:
- The TOMAHAQ technique shows promise for high-throughput quantification in proteomics.
- Interference from TMTzero reporter ions can compromise data reliability.
- A post-acquisition data correction strategy is essential for accurate TOMAHAQ implementation and unlocking its full potential.
