使用一种与 10× Genomics 单细胞 RNA-Seq 兼容的 SPAAC 无铜方法进行抗体-寡核苷酸偶联。

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

机构信息

Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore.

Molecular Engineering Laboratory, Institute of Molecular and Cell Biology (IMCB), Singapore, Singapore.

出版信息

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

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

PMID:35344168

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.

摘要

近年来，单细胞分析的多模态方法取得了进展，提供了有价值的数据点，可以补充标准流式细胞术数据。特别是，将细胞表面蛋白质组数据与基因表达分析叠加在一起是必要的进展，特别是在免疫学领域。在这里，我们描述了一种无铜点击化学方法来生成抗体-寡核苷酸复合物，并介绍了在 10× 基因组学基于液滴的单细胞 RNA-seq 工作流程中使用该方法的步骤，提供了一种有效且具有成本效益的方法来结合蛋白质组学和转录组学分析。