Friday, May 25, 2018

Chemical mediated proteolysis opens a whole new realm in middle-down proteomics!

Okay -- so -- as much as I love the concept of top-down proteomics, I don't actually do it in our lab. When a new and really difficult protein or proteomics project comes in I try to see if top-down is the solution. I really do. Like every time. With our best-suited equipment to the task (QE HF with protein mode and Fusion gen 1) I've got some size and complexity limitations that always make top-down seem ----just----this----far--- out of reach.

Conversely, I'm seeing more and more projects where trypsin or GluC or any of the other awesome enzymes in our freezer I always forget the keycode to -- just aren't right either.

Is it middle-down time??  It can't be. That only works for monoclonal antibodies, right?

No longer, thanks to this awesome paper from Kristina Srzentic et al.,!

This team details the use of a number of chemicals that can be used for middle down applications all the way down to plotting the sizes of the fragments produced and -- so important, and often overlooked --- provide a table of the mass shifts incurred by such reagents.

Okay -- at the risk of annoying my dear friends at ACS and JPR, I'm putting this table here as my civic duty to people to maybe make it just a little easier to find (ACS, please contact me directly if this is a real problem (, p.s., you're the best!)

This is just another great new tool for our utility belts for when the next weird protein (or, more likely, PTM) comes through the door!

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