Top-Down Mass Spectrometry of a Clinical Antibody Light Chain Using the Omnitrap-Orbitrap-Booster Platform

Area of Science:

• Proteomics
• Analytical Chemistry
• Biochemistry

Background:

• Top-down mass spectrometry (TD MS) is crucial for protein characterization.
• Existing TD MS platforms face challenges in fragmentation efficiency and spectral quality.
• The Omnitrap-Orbitrap-Booster (OOB) platform integrates advanced technologies to address these limitations.

Purpose of the Study:

• To evaluate the performance of the OOB mass spectrometry (MS) platform for electron capture dissociation (ECD)-based top-down (TD) MS analysis.
• To assess the OOB platform's capability for comprehensive protein characterization.
• To compare ECD-based TD MS performance on the OOB platform with established methods.

Main Methods:

• Development and integration of the Omnitrap-Orbitrap-Booster (OOB) MS platform.
• Utilizing electron capture dissociation (ECD) for tandem mass spectrometry (MS/MS) analysis.
• Performing liquid chromatography-tandem mass spectrometry (LC-MS/MS) on a P15 multiple myeloma antibody light chain across multiple precursor charge states.

Main Results:

• Single precursor charge state analysis yielded 68% sequence coverage for the P15 light chain.
• Method development using multiple precursor charge states (15+ to 19+) increased P15 sequence coverage to 93%.
• ECD-based TD MS performance on the OOB platform was comparable to electron transfer/higher-energy collision dissociation (EThcD)-based TD MS, with reduced spectral congestion.

Conclusions:

• The OOB platform significantly enhances TD MS analysis of proteins.
• Comprehensive protein characterization is achievable with optimized methods on the OOB platform.
• The OOB platform represents a powerful and efficient tool for advanced proteomics research.