Antibody-Oligonucleotide Conjugation Using a SPAAC Copper-Free Method Compatible with 10× Genomics' Single-Cell...

Area of Science:

• Immunology
• Molecular Biology
• Biotechnology

Background:

• Multimodal single-cell analyses offer valuable data complementing traditional methods like flow cytometry.
• Integrating cell-surface proteome data with gene expression analysis is crucial for advancing immunology.
• Existing methods may lack efficiency or cost-effectiveness in combining proteomic and transcriptomic data.

Purpose of the Study:

• To describe a novel copper-free click chemistry method for generating antibody-oligonucleotide complexes.
• To present a workflow for integrating this method into the 10× Genomics single-cell RNA-sequencing (scRNA-seq) platform.
• To enable efficient and cost-effective coupling of proteomic and transcriptomic analyses at the single-cell level.

Main Methods:

• Development of a copper-free click chemistry reaction for conjugating antibodies with oligonucleotide tags.
• Adaptation of the antibody-oligonucleotide complexes for use with the 10× Genomics droplet-based scRNA-seq workflow.
• Implementation of a protocol for simultaneous proteomic and transcriptomic profiling of single cells.

Main Results:

• Successful generation of antibody-oligonucleotide conjugates using copper-free click chemistry.
• Demonstrated compatibility of the method with the 10× Genomics scRNA-seq platform.
• Established a streamlined workflow for multimodal single-cell analysis, efficiently integrating proteomic and transcriptomic data.

Conclusions:

• The described copper-free click chemistry method provides an efficient and cost-effective means to couple proteomic and transcriptomic analyses in single-cell studies.
• This approach significantly advances multimodal single-cell analysis, particularly within immunological research.
• The method facilitates deeper insights into cellular heterogeneity by simultaneously measuring surface proteins and gene expression.