Wednesday, June 19, 2019

Same everything -- except buffer -- 30% increase in IDs!

I totally dig this new study's Elfseverer again....I swear, I'm not doing this to discriminate against people in the EU. Papers show up in my feed and I try to read them without looking at the author names or what journal it came from to avoid my powerful subconscious biases -- and people still submit cool things to these journals. Wait -- shit -- I may have just been emailed that I was author 84/219 on something that went to one...I should write less and read more emails....

What was I....THIS!

This team takes a look at running the same sample, gradient, instrument parameters and data processing scheme and just switches up buffer B.

Quick note: If you've got a slEasyNLC you should definitely talk to your technical support or service engineer before even considering replicating this. Maybe any nanoLC, honestly.... definitely verify that you can run buffers other than the approved ones. EasyNanos will go through seals rapidly if you use more than 80% acetonitrile as your running buffer. I think if you put 100% acetone in buffer B....

However, this brave team obviously checked with their LC manufacturer, found it was all good and swapped up the buffers on their LC and -- seriously 30% increase in HeLa IDs in what was a sub-2 hour gradient by swapping acetonitile with methanol and then acetone.

What surprises me the most, perhaps, is that the elution profile is even similar.  If at 3% acetone, all the peptides came off in one single peak, I'd be less surprised. The detector was an Orbitrap Velos running in high/low mode and the RAW files are available on ProteomeXchange here (but have yet to go live as of the time of this post's rambling).

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