High Throughput Flow Cytometry for Cell Surface Profiling.

Area of Science:

• Cellular and Molecular Biology
• Immunology
• Biomarker Discovery

Background:

• Cell surface proteins are crucial for identifying biomarkers and therapeutic targets.
• Limited experimental data exists on the surfaceome of individual cells, especially in heterogeneous samples.
• Transcriptome and surfaceome often show poor correlation, necessitating direct surface protein analysis.

Purpose of the Study:

• To develop and describe a method for high-throughput cell surface profiling using flow cytometry.
• To enable the discovery of novel cell surface markers and subpopulations.
• To provide a cost-effective, scalable approach for analyzing the surfaceome.

Main Methods:

• Utilized conventional single or multicolor flow cytometry for cell surface protein analysis.
• Developed a microarray-like screening platform for comprehensive surfaceome interrogation.
• Adapted the method for flexible antibody panel sizes, enabling customized screening.

Main Results:

• Demonstrated the capability of flow cytometry to profile cell surface proteins at single-cell resolution.
• Showcased the platform's ability to reveal unexpected markers and rare cell subpopulations.
• Generated reliable, cost-effective data from diverse sample types.

Conclusions:

• High-throughput flow cytometry offers a powerful, scalable solution for cell surface profiling.
• This approach facilitates the discovery of novel biomarkers and enhances the understanding of cellular heterogeneity.
• The method supports advanced bioinformatics analyses, such as hierarchical clustering, for deeper insights.