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Single-Cell Protein Profiling by Microdroplet Barcoding and Next-Generation Sequencing.

Samuel C Kim1,2, John R Haliburton3, Zev J Gartner4

  • 1Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, San Francisco, CA, USA. sam.kim19@gilead.com.

Methods in Molecular Biology (Clifton, N.J.)
|November 12, 2021
PubMed
Summary
This summary is machine-generated.

This study introduces DNA-barcoded antibodies for high-throughput, single-cell protein analysis. This method converts protein signals into DNA signals, enabling multiplexed protein detection alongside other single-cell profiling techniques.

Keywords:
DNA barcodingDroplet microfluidicsNext-generation sequencingProteomicsSingle-cell analysis

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Genomics

Background:

  • Single-cell analysis is crucial for understanding cellular heterogeneity.
  • Current methods for multiplex protein analysis at the single-cell level face limitations.
  • DNA barcoding offers a powerful tool for high-throughput molecular detection.

Purpose of the Study:

  • To develop a method for DNA barcoding of antibodies for single-cell protein analysis.
  • To enable sequencing-based protein quantitation by converting protein signals to DNA signals.
  • To integrate protein profiling with single-cell transcriptomics and mutational analysis.

Main Methods:

  • Preparation of DNA-barcoded antibodies targeting specific proteins.
  • Utilizing droplet microfluidics to link protein identity to cellular identity.
  • Employing next-generation sequencing for high-throughput parallel analysis of DNA barcodes.

Main Results:

  • Successful preparation of DNA-barcoded antibodies.
  • Demonstration of multiplex single-cell protein detection.
  • Compatibility of the method with single-cell transcriptomic and mutational profiling.

Conclusions:

  • DNA-barcoded antibodies provide a versatile platform for multiplex single-cell protein analysis.
  • This approach enhances the capabilities of single-cell omics studies.
  • The method facilitates comprehensive understanding of cellular states and functions.