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Spatially resolved top-down proteomics of tissue sections based on a microfluidic nanodroplet sample preparation platform

ABSTRACT: We demonstrate spatially resolved top-down proteomics (TDP) for proteoform identification and quantification from small sections of rat brain. We used the nanoPOTS (nanodroplet Processing in One pot for Trace Samples) platform, further improving the sample preparation by adding benzonase in the extraction buffer. We increased identified proteoforms from 493 to 700 in 200 cultured cells, with the largest improvements among nuclear proteins. We then performed nanoPOTS TDP on 100,000 um2 (around 200 cells) laser capture microdissectioned rat brain cortex/hypothalamus tissues. The extracted proteins were directly subjected to top down analysis without enzymatic digestion. Data was searched with TopPIC and TDPortal.

INSTRUMENT(S): Orbitrap Eclipse, Orbitrap Fusion Lumos

ORGANISM(S): Rattus Norvegicus (ncbitaxon:10116) Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Mowei Zhou

PROVIDER: MSV000089163 | MassIVE | Wed Mar 30 09:24:00 BST 2022

REPOSITORIES: MassIVE

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