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

Dominic Paul Lee; Wang Jiehao Ray; Tan Pee Mei; Shawn Hoon; Jonathan Scolnick; Gene W Yeo

doi:10.1007/978-1-0716-2160-8_6

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

Methods Mol Biol. 2022:2463:67-80. doi: 10.1007/978-1-0716-2160-8_6.

Authors

Dominic Paul Lee¹, Wang Jiehao Ray¹, Tan Pee Mei², Shawn Hoon², Jonathan Scolnick¹, Gene W Yeo^{3

4}

Affiliations

¹ Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore.
² Molecular Engineering Laboratory, Institute of Molecular and Cell Biology (IMCB), Singapore, Singapore.
³ Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore. gene.w.yeo@gmail.com.
⁴ Department of Cellular and Molecular Medicine, University of California San Diego, La Jolla, CA, USA. gene.w.yeo@gmail.com.

PMID: 35344168
DOI: 10.1007/978-1-0716-2160-8_6

Abstract

Recent advances in multimodal approaches toward single-cell analyses present valuable data points that can complement standard flow cytometry data. In particular, the overlay of cell-surface proteome data with gene expression analysis presents a necessary advancement, particularly in the field of immunology. Here we describe a copper-free click chemistry method for the generation of antibody-oligonucleotide complexes and present the steps for its employment in the context of the 10× genomics droplet-based single-cell RNA-seq workflow, providing a method for coupling proteomic and transcriptomic analyses in an efficient and cost-effect manner.

Keywords: 10× Genomics; Antibody conjugation; Multi-omics; SPAAC; Single-cell RNA-seq; Single-cell proteomics.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Antibodies
Click Chemistry / methods
Oligonucleotides* / genetics
Proteomics*
RNA-Seq

Substances

Antibodies
Oligonucleotides