Tuesday, July 28, 2015
Process DIA data directly in Proteome Discoverer 2.0 with DIA Umpire
Alright, this DIA stuff is confusing. There are methods all over the place. WiSIMDIA on the Fusion, pSMART, multiplex-DIA, and even boring old SWATH. Software-wise, there is tons of stuff out there. This weekend I processed some DIA data from a Fusion directly through Proteome Discoverer 2.0...and it looks amazing.
The data in question was ran through the DIA-Umpire to convert the data into a handy-dandy MGF file format. You can find details on the DIA-Umpire in this Nature Methods paper by Chih-Chiang Tsou et al., out of Alexey Nezvizhskii's lab. Essentially, it is a (currently) command line driven program that takes your DIA data and "deconvolutes" it down to a format that is friendly to the proteomics processing pipelines we already know and trust. How does it work? No idea. But it works, and the data looks amazing (did I say that once already?)
Here is a random high scoring PSM I grabbed. Looks pretty incredible, right? They all do. And I didn't have to change my workflow at all. I used SequestHT, target decoy and my normal basic consensus report. I ended up with a ton of IDs and really nice true FDRs at every level I set them at (PSM, peptide, and protein).
If you are interested in identifying peptides via DIA and you are a little swamped by your software options, you might want to check this out. I'm tired of learning new software interfaces -- lets put everything in Discoverer!