"Cell Surface Capture" Workflow for Label-Free Quantification of the Cell Surface Proteome

Area of Science:

• Proteomics
• Cell Biology
• Biochemistry

Background:

• Mass spectrometry-based proteomics has advanced biological system characterization.
• The cell surface proteome (surfaceome) is crucial for understanding human diseases and developing therapeutics.
• Analyzing the surfaceome is challenging due to high-abundance proteins masking low-abundance targets in cell lysates.

Purpose of the Study:

• To develop and present a detailed workflow for the accurate characterization of the cell surface proteome.
• To overcome the technical limitations in analyzing membrane and surface proteins.
• To enable precise definition of the cell surface proteome for various cell types.

Main Methods:

• A workflow for labeling cell surface proteins from cancer cells.
• Enrichment of labeled cell surface proteins from complex cell lysates.
• Sample preparation optimized for subsequent mass spectrometry analysis.

Main Results:

• A robust method for isolating and preparing cell surface proteins.
• Improved identification of cell surface proteins in cancer cells.
• Demonstration of a workflow to overcome abundance challenges in surfaceome analysis.

Conclusions:

• The presented workflow enables accurate mass spectrometry-based characterization of the cell surface proteome.
• This method facilitates the identification of potential diagnostic markers and therapeutic targets.
• The workflow addresses a critical technical barrier in surfaceome research.