TMT based LC-MS3

Highest Accuracy Workflow

Multiplexing for More Complex Samples

TMT LC-MS3 delivers the most accurate quantitative results whilst retaining good coverage in complex samples

TMT LC-MS3 is the standard method for biomarker discovery in plasma and other highly complex samples where quantitative accuracy is the most important factor. In such samples, quantitation by MS2 (MS/MS) fragmentation can become compromised and a further round of fragmentation (MS3 or MS/MS/MS) is required. Whilst using MS3 is slower and so reduces total proteome coverage, the gain in quantitative accuracy can reveal biomarkers that would be missed by other techniques. Typically, the TMT 11 or 18 plex reagents are used to allow multiplexing across study cohorts. A common study reference is included in all TMT 11 or 18 plexes to allow measurement and removal of batch effects and calculation of expression ratios.

Applications Sample Types Species Outcomes
Preclinical Drug Development Cell culture, Tissue biopsy, Xenograft Tumors Bacteria, Mammal, Insect Target Discovery / Validation
Mechanism of Action / Resistance
Clinical Drug Development Biopsy Tissue, Peripheral Fluids Human Mechanism of Action / Resistance
Biomarker Discovery Biopsy Tissue, Peripheral Fluids Human, Rodent, Other Candidate Biomarker List

TMT LC-MS3 data is processed through an integrated bioinformatics pipeline providing data integration and pre-processing (DIANA), statistical analysis (FeaST) and functional analysis (FAT) with outputs suitable for further client-led analysis and presentation to management.

The TMT LC-MS3 Workflow Offers:

  • Unbiased survey of regulated cell biology in cell lysates and tissue samples with the highest quantitative accuracy
  • Unbiased survey of regulated proteins and peptides in complex peripheral fluids with the highest quantitative accuracy
  • Combined power of TMT 11 or 18 plex reagents and Orbitrap mass spectrometers
  • Can be combined with a wide range of sample preparation techniques including fractionation and enrichment strategies
  • Standardized data outputs in Excel and ‘TSV’ files
  • Automated annotation of biological features from UniProtKB, Entrez IDs and Reactome
  • Results in 6 to 8 weeks (based on up to a 2 x 18-plex sample study)
  • Simple report highlighting key biological events and regulated features enriched in the study
  • Suitable for enhancing drug discovery, biomarker discovery, pre-clinical and clinical diagnostic assay applications