Thursday, April 9, 2015

Is this the ultimate phosphopeptide separation study?

I'm not a chemist. I assume that this fact is abundantly clear from most of these blog posts. I'm a biologist. I need this chemistry stuff (particularly methods) set out in front of me in an easily digestible format.  Then the next time some other biologist asks me how to best set up their experiment I can lead them directly to that great chemistry paper.

Some labs are very good at this side of things. Point in case, this new paper from Andrew Alpert, Otto Hudecz and Karl Mechtler.  This study starts out by very simply breaking down phosphopeptide separation chemistry in such a clear way that you'd think that this was a reference paper.  Simple and concise. At this pH, phosphopeptides stick to this thing and at this pH they don't.

Then! They go through step by step and compare every currently popular separation technique for phosphopeptide chromatography:

WAX (weak anion exchange)
SAX (strong anion exchange)
AEX (anion exchange chromatography)
ERLIC  (electrostatic repulsion-hydrophilic interaction chromatography)

Incidentally, this is essentially the title of the paper.  How clear is that?

"Hey Ben, how do you best separate phosphopeptides and why?" This is the paper that will be attached to my reply.

(The answer?  Oh yeah! It kind of depends on what phosphorylation sites you are interested in, but it looks like low pH ERLIC is going to give you the highest number of phosphopeptides if you're just going for a bulk study).

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