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Updated: Jul 6, 2026
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"Cell Surface Capture" Workflow for Label-Free Quantification of the Cell Surface Proteome
Published on: March 24, 2023
Sample Preparation Strategies for Microbial Cell Surface Proteomics: Integrating Shaving and Shotgun Approaches
Dorota Satala1, Katarzyna Kowalik1,2, Justyna Karkowska-Kuleta3
1Department of Comparative Biochemistry and Bioanalytics, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Kraków, Poland.
Advances in Experimental Medicine and Biology
|July 4, 2026
Summary
Characterizing fungal surface proteins is vital for understanding infections. Cell surface shaving enriches surface proteins for mass spectrometry, but requires careful optimization to avoid contamination and biases.
Area of Science:
- Microbiology
- Proteomics
- Biochemistry
Background:
- Pathogenic microorganisms, including fungi, possess surface proteins crucial for host interactions.
- Characterizing the fungal surface proteome is essential for developing targeted antifungal strategies.
- Current proteomic methods face challenges in obtaining pure surface-enriched samples, risking intracellular protein contamination.
Purpose of the Study:
- To discuss the challenges and strategies for optimizing sample preparation in fungal surface proteomics.
- To highlight the importance of cell surface shaving for enriching surface-exposed peptides.
- To address the need for validated methods that minimize contamination and analytical biases.
Main Methods:
- Cell surface shaving using proteolytic enzymes on intact microbial cells.
- High-resolution liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) for peptide analysis.
- Development and validation of controls to assess sample-treatment selectivity.
Main Results:
- Cell surface shaving effectively enriches for surface-exposed peptides.
- Optimization is critical to reduce sample complexity and ensure LC-MS/MS compatibility.
- Careful method development is needed to avoid matrix interferences, partial lysis, and epitope biases.
Conclusions:
- Optimized sample preparation is paramount for accurate fungal surface proteome analysis.
- Cell surface shaving offers a promising approach but requires rigorous validation.
- Addressing methodological challenges ensures reliable data for understanding fungal pathogenesis and developing therapeutics.
