Single-Cell Antibody Sequencing in Atherosclerosis Research

Ryosuke Saigusa, Christopher P. Durant, Vasantika Suryawanshi, Klaus Ley

Research output: Chapter in Book/Report/Conference proceedingChapter

2 Scopus citations

Abstract

The transcriptomic information obtained by single cell RNA sequencing (scRNA-seq) can be supplemented by information on the cell surface phenotype by using oligonucleotide-tagged monoclonal antibodies (scAb-Seq). This is of particular importance in immune cells, where the correlation between mRNA and cell surface expression is very weak. scAb-Seq is facilitated by the availability of commercial antibodies and antibody mixes. Now panels of up to 200 antibodies are available for human and mouse cells. Proteins are detected by antibodies conjugated to a tripartite DNA sequence that contains a primer for amplification and sequencing, a unique oligonucleotide that acts as an antibody barcode and a poly(dA) sequence, simultaneously detecting extension of antibody-specific DNA sequences and cDNAs in the same poly(dT)-primed reaction. For each cell, surface protein expression is captured and sequenced along with the cell’s transcriptome. Here, we list the steps needed to produce antibody sequencing data from tissue or blood cells.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages765-778
Number of pages14
DOIs
StatePublished - 2022
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume2419
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • AbSeq
  • Cellular indexing of transcriptomes and epitopes by sequencing (CITE-seq)
  • RNA expression and protein sequencing assay (REAP-seq)
  • Surface markers
  • TotalSeq

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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