Improving oligo-conjugated antibody signal in multimodal single-cell analysis

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

Improving oligo-conjugated antibody signal in multimodal single-cell analysis. / Buus, Terkild B.; Herrera, Alberto; Ivanova, Ellie; Mimitou, Eleni; Cheng, Anthony; Herati, Ramin S.; Papagiannakopoulos, Thales; Smibert, Peter; Odum, Niels; Koralov, Sergei B.

In: eLife, Vol. 10, 2021.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Buus, TB, Herrera, A, Ivanova, E, Mimitou, E, Cheng, A, Herati, RS, Papagiannakopoulos, T, Smibert, P, Odum, N & Koralov, SB 2021, 'Improving oligo-conjugated antibody signal in multimodal single-cell analysis', eLife, vol. 10. https://doi.org/10.7554/eLife.61973

APA

Buus, T. B., Herrera, A., Ivanova, E., Mimitou, E., Cheng, A., Herati, R. S., Papagiannakopoulos, T., Smibert, P., Odum, N., & Koralov, S. B. (2021). Improving oligo-conjugated antibody signal in multimodal single-cell analysis. eLife, 10. https://doi.org/10.7554/eLife.61973

Vancouver

Buus TB, Herrera A, Ivanova E, Mimitou E, Cheng A, Herati RS et al. Improving oligo-conjugated antibody signal in multimodal single-cell analysis. eLife. 2021;10. https://doi.org/10.7554/eLife.61973

Author

Buus, Terkild B. ; Herrera, Alberto ; Ivanova, Ellie ; Mimitou, Eleni ; Cheng, Anthony ; Herati, Ramin S. ; Papagiannakopoulos, Thales ; Smibert, Peter ; Odum, Niels ; Koralov, Sergei B. / Improving oligo-conjugated antibody signal in multimodal single-cell analysis. In: eLife. 2021 ; Vol. 10.

Bibtex

@article{b39898cffa5d4512baf9d9b16e837897,
title = "Improving oligo-conjugated antibody signal in multimodal single-cell analysis",
abstract = "Simultaneous measurement of surface proteins and gene expression within single cells using oligo-conjugated antibodies offers high-resolution snapshots of complex cell populations. Signal from oligo-conjugated antibodies is quantified by high-throughput sequencing and is highly scalable and sensitive. We investigated the response of oligo-conjugated antibodies towards four variables: concentration, staining volume, cell number at staining, and tissue. We find that staining with recommended antibody concentrations causes unnecessarily high background and amount of antibody used can be drastically reduced without loss of biological information. Reducing staining volume only affects antibodies targeting abundant epitopes used at low concentrations and is counteracted by reducing cell numbers. Adjusting concentrations increases signal, lowers background, and reduces costs. Background signal can account for a major fraction of total sequencing and is primarily derived from antibodies used at high concentrations. This study provides new insight into titration response and background of oligo-conjugated antibodies and offers concrete guidelines to improve such panels.",
keywords = "Background signal, CITE-seq, cytometry, ECCITE-seq, genetics, genomics, human, immunology, inflammation, oligo-conjugated antibodies, scRNA-seq",
author = "Buus, {Terkild B.} and Alberto Herrera and Ellie Ivanova and Eleni Mimitou and Anthony Cheng and Herati, {Ramin S.} and Thales Papagiannakopoulos and Peter Smibert and Niels Odum and Koralov, {Sergei B.}",
year = "2021",
doi = "10.7554/eLife.61973",
language = "English",
volume = "10",
journal = "eLife",
issn = "2050-084X",
publisher = "eLife Sciences Publications Ltd.",

}

RIS

TY - JOUR

T1 - Improving oligo-conjugated antibody signal in multimodal single-cell analysis

AU - Buus, Terkild B.

AU - Herrera, Alberto

AU - Ivanova, Ellie

AU - Mimitou, Eleni

AU - Cheng, Anthony

AU - Herati, Ramin S.

AU - Papagiannakopoulos, Thales

AU - Smibert, Peter

AU - Odum, Niels

AU - Koralov, Sergei B.

PY - 2021

Y1 - 2021

N2 - Simultaneous measurement of surface proteins and gene expression within single cells using oligo-conjugated antibodies offers high-resolution snapshots of complex cell populations. Signal from oligo-conjugated antibodies is quantified by high-throughput sequencing and is highly scalable and sensitive. We investigated the response of oligo-conjugated antibodies towards four variables: concentration, staining volume, cell number at staining, and tissue. We find that staining with recommended antibody concentrations causes unnecessarily high background and amount of antibody used can be drastically reduced without loss of biological information. Reducing staining volume only affects antibodies targeting abundant epitopes used at low concentrations and is counteracted by reducing cell numbers. Adjusting concentrations increases signal, lowers background, and reduces costs. Background signal can account for a major fraction of total sequencing and is primarily derived from antibodies used at high concentrations. This study provides new insight into titration response and background of oligo-conjugated antibodies and offers concrete guidelines to improve such panels.

AB - Simultaneous measurement of surface proteins and gene expression within single cells using oligo-conjugated antibodies offers high-resolution snapshots of complex cell populations. Signal from oligo-conjugated antibodies is quantified by high-throughput sequencing and is highly scalable and sensitive. We investigated the response of oligo-conjugated antibodies towards four variables: concentration, staining volume, cell number at staining, and tissue. We find that staining with recommended antibody concentrations causes unnecessarily high background and amount of antibody used can be drastically reduced without loss of biological information. Reducing staining volume only affects antibodies targeting abundant epitopes used at low concentrations and is counteracted by reducing cell numbers. Adjusting concentrations increases signal, lowers background, and reduces costs. Background signal can account for a major fraction of total sequencing and is primarily derived from antibodies used at high concentrations. This study provides new insight into titration response and background of oligo-conjugated antibodies and offers concrete guidelines to improve such panels.

KW - Background signal

KW - CITE-seq

KW - cytometry

KW - ECCITE-seq

KW - genetics

KW - genomics

KW - human

KW - immunology

KW - inflammation

KW - oligo-conjugated antibodies

KW - scRNA-seq

U2 - 10.7554/eLife.61973

DO - 10.7554/eLife.61973

M3 - Journal article

C2 - 33861199

AN - SCOPUS:85104390686

VL - 10

JO - eLife

JF - eLife

SN - 2050-084X

ER -

ID: 261053650