SPS MS3 Workflows – Maximize quantitative accuracy and precision

Maximize quantitative accuracy and precision

The synchronous precursor selection (SPS)-based MS3 technology on the Orbitrap Fusion Tribrid systems provide a unique, unmatched capability to accurately measure the most subtle changes in low-abundance proteins. This workflow allows for simultaneous quantitative analysis of ten samples, with significantly improved accuracy achieved by reducing the co-isolation of tagged interferences. The increased sensitivity and ion transmission boosts the number of quantifiable peptides present at low levels, enabling scientists to perform comprehensive proteome-wide quantitation faster and more accurately than ever before.

Featured TMT multiplexing technologies

Scientists pushing the limits of quantitation and protein characterization require more expansive and in-depth analytical information. The Orbitrap Fusion Lumos is the newest Tribrid mass spectrometer providing the highest sensitivity, highest selectivity and lowest detection limit. It was uniquely developed to expand the Tribrid performance in advanced -omics and biopharma workflows, including low level quantitation, data independent and top down analyses.

Isobaric chemical tags represent a fast, unbiased, and sensitive method to quantify almost all proteins in a biological sample. The TMT10plex Label Reagents share an identical structure with TMTzero, TMTduplex, and TMTsixplex Reagents but contain different numbers and combinations of 13C and 15N isotopes in the mass reporter.


What is Synchronous Precursor Selection?

The SPS method begins with selection of the parent ion in the MS scan, followed by its isolation in the quadrupole and fragmentation by collisionally induced dissociation (CID) in the ion trap. Following fragmentation, SPS enables simultaneous isolation of up to 20 MS2 fragment ions. A select group of MS2 fragment ions are then transferred back into the ion routing multipole (IRM) where they undergo higher energy collisional dissociation (HCD) fragmentation, with the MS3 fragments then detected in the Orbitrap analyzer.

The use of SPS dramatically increases the reporter ion signal intensity and improves the ratio accuracy, due to improved counting statistics, leading to a significant increase in the number of quantified peptides.


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Groundbreaking biological research enabled by technological advances in quantitative proteomics

There has been a recent and significant increase in groundbreaking biological research, relying on the TMT SPS MS3 method. The citations below are a small representation, indicating how cell biologists have capitalized on the use of this new method to help answer complex biological questions. The average impact factor for all published articles is approximately 15, about twice the impact factor of publishing in other reputable journals.


Featured citations


Mass spectrometry overview — An introduction to MS – sample prep to MS analysis — Learn more