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A Comparative Analysis of Data Analysis Tools for Data-Independent Acquisition Mass Spectrometry
Fangfei Zhang1, Weigang Ge2, Lingling Huang2
1Center for Intelligent Proteomics, Westlake Laboratory of Life Sciences and Biomedicine, Hangzhou, Zhejiang Province, China; Key Laboratory of Structural Biology of Zhejiang Province, School of Life Sciences, Westlake University, Hangzhou, Zhejiang Province, China.
Data-independent acquisition (DIA) mass spectrometry generates reproducible proteome data. Library-free DIA analysis tools show promise, but comprehensive spectral libraries remain beneficial for DIA-mass spectrometry.
Area of Science:
- Proteomics
- Mass Spectrometry
- Bioinformatics
Background:
- Data-independent acquisition (DIA) mass spectrometry is a powerful technique for reproducible proteome analysis.
- The complexity of DIA data necessitates diverse analytical tools, leading to the development of various software solutions.
- Evaluating these tools is crucial for optimizing proteomic data processing and interpretation.
Purpose of the Study:
- To assess and compare the performance of five leading DIA data analysis tools: OpenSWATH, EncyclopeDIA, Skyline, DIA-NN, and Spectronaut.
- To evaluate both library-based and library-free DIA analysis strategies across different mass spectrometry platforms.
- To provide guidance on selecting appropriate DIA analysis tools for researchers.
Main Methods:
- Utilized six DIA datasets generated from TripleTOF, Orbitrap, and TimsTOF Pro instruments.
- Compared tool performance using identification and quantification metrics.
- Analyzed shared and unique identifications across different software tools.
Main Results:
- Library-free DIA analysis approaches demonstrated superior performance when spectral libraries lacked comprehensiveness.
- Library-based methods still provided advantages in many DIA analysis scenarios, especially with comprehensive libraries.
- Significant variations in performance were observed among the evaluated DIA tools.
Conclusions:
- The choice between library-based and library-free DIA analysis depends on spectral library quality and experimental goals.
- Comprehensive spectral libraries remain valuable for maximizing proteomic data depth and accuracy in DIA-mass spectrometry.
- This comparative study offers essential insights for users of DIA-mass spectrometry technology.
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