Top-Down Proteoform Analysis by 2D MS with Quadrupolar Detection

Area of Science:

• Analytical Chemistry
• Spectrometry
• Biochemistry

Background:

• Two-dimensional mass spectrometry (2D MS) is a powerful technique for analyzing complex samples.
• Traditional 2D MS on FT-ICR MS relies on ion modulation for correlating precursor and fragment ions.
• Ion isolation is typically required, limiting multiplexing capabilities.

Purpose of the Study:

• To introduce quadrupolar detection for 2D MS within a dynamically harmonized ICR cell.
• To explore the advantages of quadrupolar detection in enhancing 2D MS capabilities.
• To develop and apply novel workflows for biomolecule analysis and quantification using this new method.

Main Methods:

• Implementation of 2D MS with quadrupolar detection in a harmonized ICR cell.
• Adaptation of data processing techniques for accurate frequency-to-mass conversion.
• Application to top-down analysis of covalently labeled ubiquitin using ECD fragmentation.

Main Results:

• Successful demonstration of 2D MS with quadrupolar detection in a harmonized ICR cell.
• Established advantages of quadrupolar detection for comprehensive fragmentation analysis.
• Developed a workflow for label-free relative quantification of biomolecule isoforms.

Conclusions:

• Quadrupolar detection significantly enhances 2D MS capabilities by eliminating the need for ion isolation.
• This method provides a robust platform for detailed biomolecule characterization.
• The developed workflow facilitates label-free quantification of biomolecule isoforms, advancing proteomic studies.