6-Plex Tandem Phosphorus Tags (TPT) for Accurate Quantitative Proteomics

Area of Science:

• Proteomics
• Mass Spectrometry
• Chemical Biology

Background:

• Isobaric chemical labeling is crucial for high-throughput quantitative proteomics.
• Existing reagents are costly and have sensitivity limitations for trace protein detection.

Purpose of the Study:

• To develop novel 6-plex tandem phosphorus tags (TPT) for enhanced multiplexing capacity in quantitative proteomics.
• To improve cost-effectiveness and sensitivity in mass spectrometry-based proteomic analyses.

Main Methods:

• Development of 6-plex tandem phosphorus tags (TPT) incorporating high isotope enrichment of 18O.
• Utilizing phosphoramidate reporter ions with ~1 Da mass difference for detection.
• Application in high-resolution ESI-MS/MS analysis of HeLa cell tryptic peptides.

Main Results:

• Achieved a 3-fold increase in multiplexing capacity compared to previous 2-plex reagents.
• Demonstrated ultrasensitive and accurate proteome quantification.
• Confirmed high labeling efficiency for large-scale quantitative proteomics.

Conclusions:

• The novel 6-plex TPT reagents facilitate cost-effective, large-scale, and accurate quantitative proteomics.
• This advancement offers improved sensitivity and multiplexing for mass spectrometry applications.
• TPT reagents represent a significant improvement for ultrasensitive proteome quantification.