Saturday, November 25, 2017
TAFT -- Fast, reproducible phosphopeptide enrichment AND fractionation on a stage tip!
If TAFT works as well as these researchers claim in this new study, this could be a game changer!
They directly enrich phosphopeptides and then do high ph reversed phase fractionation -- all in the same tube/tip and it's ready to go!
Did the number of phosphopeptides they identify break records set by techniques like SCX or high pH RP fraction collection followed by TiO2/IMAC enrichment? No. It's not bad, but we have seen better.
However -- 4 hours from start to finish! Compare that to most of the deep phopsphoproteomics studies we've seen -- they point out several and you're looking at a full day to prep the samples and then up to 48 hours of instrument run time. 3 hours vs. a long week?!?
If you use TAFT you could use the other 4.5 days of your work week to try and make sense of the 14,000 phosphopeptides you identified.
On the method details -- they do use an Orbitrap Fusion for their analysis. It appears to be all HCD high resolution MS/MS on the mass spec side for this study and the data processing is in MaxQuant / Perseus.
The reproducibility of the TAFT protocol, run to run, looks pretty spectacular as well. One of the benefits of keeping everything simple and and all in one tube!