Pushing the leading edge in next generation quantitative proteomics
The ability to increase the number of proteomic samples that can be compared quantitatively in a single LC-MS run is highly desirable for analytical throughput. This also enables assessment of multiple experimental conditions and leads to a decrease in missing values among replicates. We have designed a new tandem mass tag set that can be used to quantify up to sixteen protein concurrently. The new tag contains nine stable isotopes and a novel proline-based reporter with the same elemental composition and mass as the TMT reporter ion. Compared to TMT reagents, TMTpro reagents have the same peptide labeling efficiency, but require slightly lower MS collision energy for reporter ion fragmentation and modifications of LC-MS gradients for optimal analysis of the labeled samples.
We also demonstrate that TMTpro 16plex samples have similar peptide/protein identification rates compared to TMT 11plex samples using a new standardized sample preparation workflow that reduces processing time from intact cells to cleaned-up peptides under 3.5 hours. This presentation will provide a technical overview of these new reagents in the context of experimental design, sample preparation, LC-MS analysis and data processing.
Who should attend:
- Director, PIs & Managers of Proteomics labs
- Director, PIs & Managers of Biopharma labs
- Current users of Q Exactive technology & QTOF MS analyzing peptides & proteins
What you will learn:
- Relative quantitation using new tandem mass tags to quantify up to 16 proteins
- Multiplexed proteomics workflow
- Experimental design
- Sample preparation
- Data processing
Presenter: Sergei I. Snovida, Ph.D. (R&D Scientist III, Protein Analysis Reagents, Thermo Fisher Scientific)
Sergei Snovida has developed many of the products for MS, including TMT reagents, instrument calibration solutions, and various sample preparation kits for proteomics experiments. Prior to obtaining his Ph.D. in Analytical Chemistry from the University of Manitoba (Canada), he worked as an analytical QC scientist at several pharma and industrial chemical companies. Sergei postdoc(ed) at Academia Sinica (Taiwan) where he worked on the development of new MS techniques for glycomics and glycoproteomics workflows. His professional interests include separations, peptides & carbohydrates, experimental design, and analytical method development and optimization in general.
