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Published on: May 1, 2017
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New Set of Isobaric Labeling Reagents for Quantitative 16Plex Proteomics.
Xiaolian Ning1,2, Qidan Li2, Jin Zi2,3
1College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049, China.
Analytical Chemistry
|March 23, 2023
Summary
Researchers developed new 16-plex isobaric tags (IBT-16plex) for improved peptide labeling. This novel reagent enhances proteome analysis by increasing identified peptides and proteins while minimizing signal suppression.
Area of Science:
- Proteomics
- Mass Spectrometry
- Chemical Biology
Background:
- Isobaric tags are crucial for relative quantitative proteome analysis across multiple sample groups.
- Advancements in isobaric reagents aim to increase multiplexing capabilities and reduce reporter ion signal suppression.
Purpose of the Study:
- To develop a novel set of 16-plex isobaric tags (IBT-16plex) with improved performance over existing reagents.
- To evaluate the efficiency and effectiveness of IBT-16plex in peptide labeling and proteome quantification.
Main Methods:
- Redesigning the chemical structure of deuterium isobaric amine-reactive tags.
- Optimizing the synthetic pathway for the new 16-plex isobaric tags.
- Comparing IBT-16plex performance against TMTpro-16plex using HeLa and MCF7 cell proteomes.
Main Results:
- IBT-16plex achieved near-complete peptide labeling within 90 minutes with comparable MS/MS signals.
- Significantly increased peptide (14.8%) and protein (8.6%) identification compared to TMTpro-16plex in HeLa cells.
- Demonstrated reduced suppression of peptide abundance differences and a wider dynamic range compared to TMTpro-16plex.
Conclusions:
- IBT-16plex offers enhanced proteome analysis capabilities, identifying more peptides and proteins.
- The novel reagent is effective in quantifying protein responses, as shown in MCF7 cells under varying glucose conditions.
- IBT-16plex represents a significant advancement for multiplexed quantitative proteomics.
