Resampling and deconvolution of linear time-of-flight records for enhanced protein profiling.

Area of Science:

• Analytical Chemistry
• Biochemistry
• Spectroscopy

Background:

• Time-of-flight (TOF) mass spectrometry is crucial for analyzing complex biological samples like protein mass spectra.
• Existing methods for TOF spectral analysis face challenges with peak resolution and sensitivity across the full mass range.
• Automated data analysis and peak detection in dense TOF spectra remain an area for improvement.

Purpose of the Study:

• To develop and validate a novel peak deconvolution strategy for time-of-flight (TOF) mass spectra.
• To enhance spectral resolution and sensitivity across the entire mass range of TOF instruments.
• To facilitate automated data analysis and peak detection in complex TOF spectra.

Main Methods:

• Developed a peak deconvolution strategy applicable to the full mass range of TOF spectra.
• Resampled spectra to create a time series with equal peak widths for consistent deconvolution.
• Applied deconvolution filters to protein mass spectra from blood serum and cell lysates acquired on three TOF instruments.

Main Results:

• Achieved a 40% improvement in spectral resolution across the full linear TOF record up to m/z 150,000.
• Enhanced experimental sensitivity, enabling better detection of spectral structures.
• Successfully resolved multiply charged ions, matrix adducts, and post-translational protein modifications.

Conclusions:

• The developed peak deconvolution strategy significantly improves TOF mass spectral analysis.
• This approach offers enhanced resolution and sensitivity, particularly beneficial for dense spectra.
• The method is well-suited for automated data analysis and peak detection in TOF mass spectrometry.