EVOSEP ONE

Importance of the Topic

The rapid and reproducible analysis of complex proteomes is critical in clinical omics research, where standardization, high throughput and inter-laboratory comparability are essential. Evosep One addresses these needs by combining robust chromatography, simplified sample handling and preset methods to accelerate biomarker discovery and routine diagnostics.

Objectives and Study Overview

This document presents the design, performance and application of the Evosep One system, a turnkey platform for gradient delivery, sample preparation and nanoflow chromatography. Key aims include:

• Demonstrating standardized methods covering 30 to 500 samples per day
• Evaluating reproducibility and robustness across runs and storage times
• Highlighting ease of integration with mass spectrometers

Methodology and Instrumentation

The Evosep One employs a two-stage pumping scheme: a low-pressure gradient generator and a high-pressure delivery module.

• Gradient methods: Six standard workflows (500/300/200/100/60/30 samples per day) with gradient lengths from 2.2 to 44 minutes and flow rates of 0.5–4.0 µl/min.
• Evotip sample traps: Disposable C18 cartridges for offline loading, desalting and storage (stable up to 35 days).
• Columns: Endurance (4–15 cm × 150 µm, 1.9 µm) and Performance (8–15 cm × 100–150 µm, 1.5–3 µm) for sharp peaks (FWHM 2.85–5.11 s).
• Nanoflow Whisper methods: Specialized low-volume (100 nl/min) workflows for single-cell and high-sensitivity proteomics.
• Integration: Zero dead-volume connectors and native drivers for Thermo, Bruker, SCIEX, Agilent systems.

Main Results and Discussion

Evosep One delivers high reproducibility and low variability across hundreds of injections:

• Run-to-run cycle times below one second overhead.
• Coefficient of variation ~6% for peptide peak areas over 96 injections (100 samples/day method).
• Stable peptide identifications from 1–50 ng HeLa digests with CVs under 10% after up to 35 days on Evotips.
• Chromatographic performance: FWHM as low as 2.85 s and 4σ peak widths under 10 s, ensuring high resolution and sensitivity.

Benefits and Practical Applications

Evosep One offers:

• High throughput without sacrificing chromatographic quality.
• Standardized workflows for reproducible inter-laboratory studies.
• Streamlined sample preparation reducing hands-on time and contamination risk.
• Flexible methods for deep proteome coverage, QA/QC and clinical applications.

Future Trends and Applications

Emerging directions include:

• Single-cell proteomics with nanoflow Whisper™ gradients.
• Extended and high-organic methods for specialized applications.
• Automated, remote-controlled operation for decentralized laboratories.
• Enhanced integration with AI-driven data analysis pipelines.

Conclusion

Evosep One provides a unified, robust solution for clinical-scale proteomics, balancing speed, sensitivity and reproducibility. Its standardized methods and simplified workflows enable large-scale studies, from biomarker discovery to routine diagnostics.

Used Instrumentation

• Evosep One separation system with dual-stage pumps
• Evotip disposable sample traps
• Endurance and Performance chromatographic columns
• Nanoflow Whisper™ gradient module
• Zero dead-volume connectors and native MS drivers

References

No specific literature references provided in the source document.