Low-flow LC-MS proteomics research. Chromatography consumables reference guide

Importance of the Topic

Low-flow liquid chromatography–mass spectrometry (LC-MS) plays a pivotal role in modern proteomics by delivering exceptional sensitivity and resolution from minimal sample volumes. Innovations in column chemistries, connection technologies, and sample preparation are driving breakthroughs in biomarker discovery, therapeutic monitoring, and fundamental protein research.

Objectives and Overview

This summary reviews Thermo Fisher Scientific's portfolio of low-flow chromatography consumables and instruments, emphasizing high-pressure nano, capillary, and micro-flow LC systems. The goal is to highlight how integrated solutions—from columns and fittings to autosampler vials and sample prep kits—enable robust, high-throughput proteome analyses.

Methodology and Instrumentation

The platform combines the Vanquish Neo UHPLC system with state-of-the-art columns, connectors, and mass spectrometers. Key components include:

• PepMap Neo columns (1500 bar rating) in EASY-Spray or Double nanoViper formats
• MAbPac RP capillary columns for intact protein separations
• Plug-and-spray EASY-Spray and tool-free Viper/nanoViper fittings
• Thermo Scientific Orbitrap Exploris 480 and TSQ Altis mass spectrometers
• SMART Digest pepsin and trypsin kits, automated on KingFisher systems
• Certified autosampler vials, closures, and 96-well plates for low-flow workflows

Main Results and Discussion

Benchmarks demonstrate:

• Single-shot nanoLC-MS bottom-up proteomics with deep proteome coverage using 75 μm × 75 cm PepMap Neo columns
• Ultra-robust micro-flow LC-MS/MS for targeted quantitation over thousands of runs
• Sensitive top-down analysis of therapeutic antibodies and subunits with MAbPac RP capillaries
• Consistent performance across 38,000 micro-flow proteome runs, illustrating system durability
• Rapid, reproducible digestion workflows in under 5 minutes using SMART Digest kits

Benefits and Practical Applications

These integrated solutions deliver:

• Enhanced sensitivity and reduced dispersion for trace-level protein detection
• High throughput and long-term robustness for large-scale proteomics projects
• Simplified setup with plug-and-play fittings and automated sample prep
• Versatility across bottom-up and top-down proteomics, biomarker discovery, and QC of biotherapeutics

Future Trends and Opportunities

Advances are expected in higher pressure-tolerant columns, further miniaturization, AI-driven method optimization, and expanded capabilities for single-cell proteomics. Integration with emerging mass spectrometry platforms will continue to enhance throughput and data quality.

Conclusion

Thermo Fisher Scientific’s low-flow LC-MS consumables and Vanquish Neo system provide a comprehensive, high-performance toolkit for modern proteomics. By combining robust instrumentation, advanced column chemistries, and efficient sample handling, laboratories can achieve new levels of sensitivity, reproducibility, and throughput.

Instrumentation Used

• Vanquish Neo UHPLC system
• Thermo Scientific Orbitrap Exploris 480 Mass Spectrometer
• Thermo Scientific TSQ Altis Triple Quadrupole Mass Spectrometer
• PepMap Neo EASY-Spray and Double nanoViper columns
• MAbPac RP capillary columns
• Thermo Scientific SMART Digest Pepsin and Trypsin Kits
• Thermo Scientific KingFisher System
• Certified autosampler vials, caps, and 96-well plates