Mass defect-based carbonyl activated tags (mdCATs) for multiplex data-independent acquisition proteome quantification

Area of Science:

• Proteomics and Mass Spectrometry
• Biomarker Discovery
• Translational Medicine

Background:

• Mass spectrometry (MS) quantification is crucial in biology and medicine.
• Data-independent acquisition (DIA) offers advantages but struggles with multiplexing due to labeling limitations.
• Existing reagents like TMT and iTRAQ are not ideal for DIA multiplexing.

Purpose of the Study:

• To develop a novel multiplex quantitative reagent for DIA.
• To enable parallel quantification of multiple samples within a single DIA injection.
• To overcome limitations of current DIA labeling strategies.

Main Methods:

• Design and synthesis of a novel mass-defect-based carbonyl activated tag (mdCAT).
• Integration of mdCATs with the DIA strategy for proteomic quantification.
• Evaluation of mdCAT performance in parallel quantification of eight samples.

Main Results:

• Successfully developed mdCATs enabling parallel quantification of eight samples with DIA.
• mdCATs eliminate spectral complexity due to minuscule mass differences.
• Achieved broad proteomic coverage, good reproducibility, and accuracy in DIA experiments.

Conclusions:

• mdCATs significantly advance DIA capabilities for multiplexed quantitative analysis.
• This technology opens new avenues for clinical sample investigation and biomarker discovery.
• The study pushes DIA towards unprecedented multiplexing and improved diagnostic biomarker identification.