Wednesday, March 23, 2022

Toward zero variance in proteomics sample preparation!


Wooohooo! 

More automation, please! 


This group demonstrates the use of an Agilent Bravo positive pressure system for sample prep of mouse heart chunks (lysate is obtained via Barocycler, I think) and then breaking out these things:  30 kDa AcroPrep Omega filter membrane plate

Lot of people are working on these things (we're working on automating 96-well S-Traps and should be up shortly!) but this group went with FASP (filter-aided sample preparation) and the results look impressively consistent. 

FASP does take a while, and I bet you can't add too much positive pressure without breaking them. Interestingly, this group quantifies their protein using amino acid analysis which probably only sounds difficult to me because I haven't done it, but -- again -- the results look fantastic. 

I figured that a big advantage over my S-Trap system would be cost savings, but there isn't as big of a gap as I'd guessed. I think we pay $350/96-well S-Trap plate. So more like $4/sample vs $3/sample. This would obviously add up over big cohorts, but it isn't something that would make me consider changing my current plans. 


In the end, if a robot is doing the pipetting in exactly the right volumes at exactly the right times, proteomics and science wins. If every study had the level of reproducibility across the board as this group shows in these impressive plots, LCMS proteomics as a field shakes off one of the biggest criticisms the outside world has historically thrown at us and we can push forward to the next challenge! Everyone wins. 

1 comment:

  1. Hi Ben, I am happy that you liked the study of my colleague :). Just a short comment, the Resolvex positive pressure device is actually from TECAN, not from Agilent.

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