A High-Sensitivity and High-Throughput LC-MS Workflow for Low-Input and Single-Cell Proteomics

A scpLASP LC-MS workflow for HeLa digests increases mean identified protein groups to 7,409 at 250 pg using Orbitrap Astral Zoom with optimized mobile phases and 5.5-min gradients.

• Why it matters: Single-cell proteomics is limited by protein scarcity and sample loss, requiring both low-loss preparation and highly sensitive, high-throughput LC-MS analysis.
• What they did: The authors optimized peptide reconstitution (0.01% DDM + 0.01% LMNG) and LC mobile phases (0.5% acetic acid + 1% DMSO), then shortened gradients from 11 to 5.5 min.
• The result: With the 5.5-min gradient, they achieved 135 samples/day for HEK293 Expi293F single cells (4.5 µL injections) and identified a median of 5,655 protein groups per cell.