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 11-, 18- or 35-plex reagents are used to allow multiplexing across study cohorts. A common study reference is included in all TMT 11-, 18- or 35-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-, 18- or 35-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