The Conjugation of Antibodies for the Simultaneous Detection of Surface Proteins and Transcriptome Analysis at a Single-Cell Level

Methods Mol Biol. 2020:2184:31-45. doi: 10.1007/978-1-0716-0802-9_3.

Abstract

Transcriptome analysis at a single-cell level with single-cell RNA sequencing (scRNA-seq) is a powerful method for detailed characterization of heterogeneous cell populations. Recent developments have enabled parallel analysis of both transcript and protein levels by using antibodies conjugated to barcoded oligonucleotides. These antibodies enable protein levels to be converted into nucleotide format, allowing the sequencing-based detection of both modalities at single-cell level. Here we present a simple and reliable method for conjugation of oligonucleotides with antibodies and a protocol for their use in single-cell transcriptome sequencing.

Keywords: CITE-seq; Proteomics; Transcriptome analysis; Transcriptomics; scRNA-seq.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies / genetics*
  • Cells, Cultured
  • Gene Expression Profiling / methods*
  • Humans
  • Leukocytes, Mononuclear / metabolism
  • Membrane Proteins / genetics*
  • Oligonucleotides / genetics
  • Sequence Analysis, RNA / methods
  • Single-Cell Analysis / methods*
  • Transcriptome / genetics*

Substances

  • Antibodies
  • Membrane Proteins
  • Oligonucleotides