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Developing an automation-ready high throughput chemoproteomic workflow


EMSL Project ID
60473

Abstract

We propose to develop a high-throughput (HTP) workflow for activity-based protein profiling (ABPP) or bioorthogonal non-canonical amino acid tagging (BONCAT) samples. This HTP chemoproteomic sample preparation workflow will include a fluorescence readout to rapidly assess sample quality (e.g. labeling efficiency) in all samples and facilitate the transition to a 96-well format for automated liquid handling and sample processing. All steps of ABPP sample preparation, from click chemistry, enrichment, digestion, and tandem mass tag (TMT) labeling of peptides, will be optimized for 96-well format. Development of this HTP workflow will make ABPP and other chemoproteomic approaches less labor intensive (lower cost) and more accessible to EMSL users.

Project Details

Start Date
2022-10-01
End Date
N/A
Status
Active

Team

Principal Investigator

Vivian Lin
Institution
Pacific Northwest National Laboratory

Co-Investigator(s)

Heather Olson
Institution
Environmental Molecular Sciences Laboratory

Team Members

Ryan Ziels
Institution
The University of British Columbia

Erik Hom
Institution
University of Mississippi

Stephen Callister
Institution
Pacific Northwest National Laboratory