diaPASEF

diaPASEF combines data-independent acquisition with parallel accumulation–serial fragmentation and ion mobility to increase specificity and quantitative accuracy in proteomic precursor identification.

Key Features:

• Data-independent acquisition (DIA): Cycles through predefined m/z segments to isolate and concurrently fragment populations of precursor ions.
• Parallel Accumulation–Serial Fragmentation (PASEF): Combines parallel accumulation with serial fragmentation to increase sampling efficiency of precursor ions.
• timsTOF Pro integration: Uses a trapped ion mobility device to exploit the correlation between molecular weight and ion mobility, enabling sampling of up to 100% of peptide precursor ion current across m/z and mobility dimensions.
• Ion mobility-aware data extraction: Incorporates the ion mobility dimension into targeted data extraction workflows for signal extraction and scoring.
• Improved specificity: Enhances precursor identification specificity through the additional ion mobility dimension.
• Reproducibility and quantitative accuracy: Increases reproducibility and quantitative precision in proteomics measurements.
• High sensitivity and coverage: Achieves deep proteome coverage from minimal sample amounts, reported down to 10 ng.
• Validation: Demonstrated on whole proteome digests and mixed organism samples.

Scientific Applications:

• Deep proteome profiling: Enables comprehensive proteome coverage in complex biological samples.
• Low-input proteomics: Supports sensitive analysis from very small sample amounts (as low as 10 ng).
• Complex-sample analysis: Applicable to whole proteome digests and mixed organism samples for robust identification and quantification.
• Quantitative DIA studies: Facilitates reproducible, quantitative proteomics using DIA combined with ion mobility.

Methodology:

Extends established targeted data extraction workflows by incorporating ion mobility as an additional dimension for ion signal extraction and scoring.