Research by Ye et al. out of the Olsen lab published in bioRxiv, introduces groundbreaking advancements in single-cell proteomics (SCP) using mass spectrometry. Single-cells were isolated and processed on the cellenONE before samples were prepared using the innovative proteoCHIP EVO 96. Next, samples were loaded onto the Evosep One LC utilising IonOpticks UHPLC columns before analysis on a Thermo Orbitrap Astral MS. Using this advanced workflow, researchers can now access a more comprehensive view of the proteome within individual cells, thanks to improvements in sensitivity, coverage, and reliability.
The study demonstrates a nearly lossless SCP workflow to identify over 5000 proteins and 40,000 peptides from single HeLa cells. These enhancements enable the direct detection of post-translational modifications (PTMs) and simplify the analytical process. With the potential to process up to 80 label-free SCP samples per day, this workflow sets a new standard for sensitivity and throughput, with applications spanning cellular development, disease progression, and cell type-specific marker identification.
Zilu Ye, Pierre Sabatier, Leander Hoeven, Teeradon Phlairaharn, David Hartlmayr, Fabiana Izaguirre, Anjali Seth, Hiren Joshi, Dorte Bekker-Jensen, Nicolai Bache, Jesper V Olsen