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Quantifying protein abundance on single cells using split-pool sequencing on DNA-barcoded antibodies for diagnostic
Jenny Sheng1,2, Eldad A Hod2, George Vlad2
1Integrated Program in Cellular, Molecular, and Biomedical Studies, Columbia University Irving Medical Center, New York, NY, 10032, USA.
Scientific Reports
|January 19, 2022
Summary
Researchers developed a new protein detection method for single cells using widely available next-generation sequencing (NGS). This accessible technique offers high-throughput proteomics and analyzes complex patient samples, outperforming traditional flow cytometry.
Area of Science:
- Proteomics
- Cellular Biology
- Biotechnology
Background:
- Protein abundance is crucial for understanding cell identity and state.
- Traditional methods like flow cytometry face limitations due to spectral overlap.
- Mass cytometry (CyTOF) offers high-plex detection but requires specialized instrumentation.
Purpose of the Study:
- To develop an accessible method for quantifying multiple protein targets on single cells.
- To overcome the limitations of spectral overlap and specialized equipment in protein detection.
- To provide a high-throughput proteomics solution for complex biological samples.
Main Methods:
- Developed a novel combinatorial indexing method for single-cell protein quantification.
- Utilized widely available next-generation sequencing (NGS) services.
- Assayed over two dozen target proteins simultaneously on single cells.
Main Results:
- The new method demonstrates favorable comparison to traditional flow cytometry.
- Successfully analyzed peripheral blood and bone marrow aspirates from human clinical samples.
- Identified pathogenic cellular subsets with high fidelity.
Conclusions:
- The developed combinatorial indexing method is an accessible and powerful tool for high-throughput single-cell proteomics.
- This technique is promising for interrogating complex samples, including those from leukemia patients.
- Offers a viable alternative to specialized instrumentation for multi-protein analysis.
