Higher-Throughput Proteome Profiling Enabled by Parallelized Pre-Accumulation and Optimized Ion Processing in the...

Area of Science:

• Proteomics
• Mass Spectrometry
• Systems Biology

Background:

• High-throughput proteomics is essential for biological research but limited by current mass spectrometry (MS) technology.
• Existing MS methods face challenges in speed, sensitivity, and scalability for large sample analyses.

Purpose of the Study:

• To introduce the Thermo Scientific Orbitrap Astral Zoom MS, designed to overcome current limitations in high-throughput proteomics.
• To enhance acquisition speed, ion utilization, and spectral processing for deeper proteome analysis.

Main Methods:

• Utilized the Orbitrap Astral Zoom MS with ultra-fast MS/MS scan rates up to 270 Hz.
• Implemented pre-accumulation for enhanced ion utilization.
• Analyzed human cell lysate for proteome depth and throughput assessment.

Main Results:

• Identified approximately 100,000 unique peptides and over 8,400 proteins in a single 300 samples-per-day (SPD) analysis.
• Achieved near-complete proteome coverage (>12,000 proteins) in 2.7 hours, a 40% reduction in analysis time.
• Demonstrated ultra-high-throughput capabilities, identifying >7,000 proteins in a 500 SPD method with high reproducibility (Pairwise Pearson correlations >0.99).

Conclusions:

• The Orbitrap Astral Zoom MS offers significant advancements in speed, sensitivity, and scalability for proteomics.
• This technology enables routine large-scale proteomics, supporting applications in clinical research and systems biology.
• The instrument represents a major step forward for high-throughput proteome analysis.