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Single-Cell Antibody Sequencing in Atherosclerosis Research

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Atherosclerosis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2419))

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.

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Acknowledgments

This study was supported by Uehara Memorial foundation fellowship to R.S., NIH HL 136275, 145241, and 148094 to K.L.

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Authors and Affiliations

  1. La Jolla Institute for Immunology, La Jolla, CA, USA

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

Authors
  1. Ryosuke Saigusa
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  2. Christopher P. Durant
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  3. Vasantika Suryawanshi
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  4. Klaus Ley
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Corresponding author

Correspondence to Klaus Ley .

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Editors and Affiliations

  1. School of Biosciences, Cardiff University, Cardiff, UK

    Dipak Ramji

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© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Saigusa, R., Durant, C.P., Suryawanshi, V., Ley, K. (2022). Single-Cell Antibody Sequencing in Atherosclerosis Research. In: Ramji, D. (eds) Atherosclerosis. Methods in Molecular Biology, vol 2419. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1924-7_46

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  • DOI: https://doi.org/10.1007/978-1-0716-1924-7_46

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1923-0

  • Online ISBN: 978-1-0716-1924-7

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