Mass spectrometry (MS)-based proteomics is an increasingly powerful analysis method that allows identification and quantitation of thousands of proteins simultaneously. MS methods used to analyze proteins fall into one of two categories: bottom-up proteomics, where proteins are first digested into peptides followed by analysis with the mass spectrometer or top-down proteomics, that analyzes the intact protein. The bottom-up approach is the more prevalent of the two methods given the current upper size limit for analysis of intact proteins using top-down proteomics. The last few decades have seen the development of different MS quantitation strategies for the bottom-up approach for both discovery and targeted proteomics. The discovery quantitation methods include label-free, metabolic labeling using heavy isotopes, and chemical labeling using isobaric tags. While each of these bottom-up methods has advantages and limitations, isobaric labeling using tandem mass tags (TMT) offers the advantage of a high degree of multiplexing without compromising protein identification and quantitation.
The past few years have also seen major advances in TMT technology, both in the level of multiplexing and in the integration of TMT tags as a method for quantitation in various areas of research. The standard tandem mass tags reagents were first developed as a 6-plex that enabled multiplex relative quantitation using high-resolution MS/MS for samples prepared from cells, tissues, or biological fluids. The level of multiplexing was then extended to a 10-plex and then an 11-plex which allowed for concurrent MS analysis of up to 11 samples. In 2019, the next generation of tandem mass tags also called TMTpro reagents were launched to enable multiplex relative quantitation of up to 16 samples. These new TMTpro reagents are similar in design to the standard tandem mass tags but differ in structure without compromising labeling efficiency and quantitative precision. Introduced earlier this year, the TMTpro 18-plex pushes the boundaries of sample multiplexing to allow for the simultaneous global protein profiling of 18 samples by LC-MS/MS analysis.
In addition to the development of these higher multiplex tandem mass tags, several TMT based protein-profiling methods have been developed and employed successfully across a wide variety of applications. These advanced applications include mapping cell-wide distribution of the proteome (hyperLOPIT), characterizing site-specific post-translational modifications (SL-TMT for phosphorylation, UbiFast for ubiquitination), profiling differences in large scale plasma studies (AutoMP3), monitoring global protein translation (mePROD), analyzing protein degradation and synthesis (mPDP), determining thermal stability and solubility of proteins, assessing protein-protein interactions using cleavable cross-linkers, multiplexing 2D methods for targeted proteomics (TOMAHAQ) and sequencing single-cell proteomes (SCeptre).
“Working in Tandem - 1st Annual TMT Symposium” is the first of a kind virtual event that brings together researchers who are utilizing TMT reagents to innovate and expand our knowledge of protein expression, modifications, localization and structure. Hear about the history and the progress of TMT technology from key leaders on their research including their latest TMT labeling reagent workflows, methods and data. Learn from the experts about best practices, tips and tricks, and how to overcome challenges that one may encounter during a TMT multiplexing experiment. At the end of each day’s program, join the experts in a live panel discussion about the day’s topics as they share insights about the latest innovations and address questions submitted during the presentations.