TAG-TMTpro, a Hyperplexing Quantitative Approach for High-Throughput Proteomic Studies.

Area of Science:

• Proteomics
• Analytical Chemistry
• Biochemistry

Background:

• Isobaric labeling techniques, such as TMTpro, are crucial for multiplexed quantitative proteomics.
• Current methods allow up to 18-plex quantification, but higher throughput is needed for comprehensive studies.

Purpose of the Study:

• To develop a novel approach to significantly increase the multiplexing capacity of TMTpro labeling.
• To establish a versatile quantitative strategy for high-throughput proteomic analyses.

Main Methods:

• Introduction of Ala or Gly residues to peptides before TMTpro labeling (TAG-TMTpro approach).
• Optimization of labeling, side-product removal, and deprotection steps.
• Validation using *E. coli* and HeLa cell lysates for identification and quantification.

Main Results:

• The TAG-TMTpro approach successfully tripled the quantitative capacity of TMTpro reagents.
• Demonstrated good identification reproducibility and reliable quantitative accuracy.
• Validated performance in complex biological samples.

Conclusions:

• TAG-TMTpro is a novel and effective method for enhancing multiplexing in TMTpro-based proteomics.
• This approach significantly advances high-throughput quantitative proteomic studies.