Cell surface capturing technologies for the surfaceome discovery of hepatocytes

Area of Science:

• Cell Biology
• Proteomics
• Biochemistry

Background:

• Cell surface proteins (surfaceome) mediate cellular interactions and environmental sensing.
• Surfaceome markers are vital for cell phenotyping, developmental staging, and disease diagnostics.
• Cell surface proteins are promising blood biomarkers due to their potential secretion or shedding.

Purpose of the Study:

• To address limitations in identifying the complete cell surface proteome.
• To present a detailed protocol for Cell Surface Capturing (CSC) technology and its variants.
• To demonstrate the utility of CSC technologies for a systems biology view of the surfaceome.

Main Methods:

• Utilized mass spectrometry-based Cell Surface Capturing (CSC) technology.
• Employed variants like Glyco-CSC, Cys-Glyco-CSC, and Lys-CSC for selective enrichment.
• Developed a step-by-step protocol applicable to both soluble and adherent cells.

Main Results:

• CSC technologies overcome limitations of antibody-based detection for cell surface proteins.
• The methods enable selective enrichment and identification of N-glycosylated or lysine-exposed proteins.
• Application to Hepa1-6 cells showcased the complementary nature of CSC technologies for surfaceome analysis.

Conclusions:

• CSC technologies provide a powerful approach for comprehensive cell surface proteome identification.
• These methods enhance the discovery of potential biomarkers from the cell surface.
• The technology facilitates a systems biology perspective on the dynamic cell surface environment.