Cargando…

Abseq: Ultrahigh-throughput single cell protein profiling with droplet microfluidic barcoding

Proteins are the primary effectors of cellular function, including cellular metabolism, structural dynamics, and information processing. However, quantitative characterization of proteins at the single-cell level is challenging due to the tiny amount of protein available. Here, we present Abseq, a m...

Descripción completa

Detalles Bibliográficos
Autores principales: Shahi, Payam, Kim, Samuel C., Haliburton, John R., Gartner, Zev J., Abate, Adam R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5349531/
https://www.ncbi.nlm.nih.gov/pubmed/28290550
http://dx.doi.org/10.1038/srep44447
_version_ 1782514485714485248
author Shahi, Payam
Kim, Samuel C.
Haliburton, John R.
Gartner, Zev J.
Abate, Adam R.
author_facet Shahi, Payam
Kim, Samuel C.
Haliburton, John R.
Gartner, Zev J.
Abate, Adam R.
author_sort Shahi, Payam
collection PubMed
description Proteins are the primary effectors of cellular function, including cellular metabolism, structural dynamics, and information processing. However, quantitative characterization of proteins at the single-cell level is challenging due to the tiny amount of protein available. Here, we present Abseq, a method to detect and quantitate proteins in single cells at ultrahigh throughput. Like flow and mass cytometry, Abseq uses specific antibodies to detect epitopes of interest; however, unlike these methods, antibodies are labeled with sequence tags that can be read out with microfluidic barcoding and DNA sequencing. We demonstrate this novel approach by characterizing surface proteins of different cell types at the single-cell level and distinguishing between the cells by their protein expression profiles. DNA-tagged antibodies provide multiple advantages for profiling proteins in single cells, including the ability to amplify low-abundance tags to make them detectable with sequencing, to use molecular indices for quantitative results, and essentially limitless multiplexing.
format Online
Article
Text
id pubmed-5349531
institution National Center for Biotechnology Information
language English
publishDate 2017
publisher Nature Publishing Group
record_format MEDLINE/PubMed
spelling pubmed-53495312017-03-17 Abseq: Ultrahigh-throughput single cell protein profiling with droplet microfluidic barcoding Shahi, Payam Kim, Samuel C. Haliburton, John R. Gartner, Zev J. Abate, Adam R. Sci Rep Article Proteins are the primary effectors of cellular function, including cellular metabolism, structural dynamics, and information processing. However, quantitative characterization of proteins at the single-cell level is challenging due to the tiny amount of protein available. Here, we present Abseq, a method to detect and quantitate proteins in single cells at ultrahigh throughput. Like flow and mass cytometry, Abseq uses specific antibodies to detect epitopes of interest; however, unlike these methods, antibodies are labeled with sequence tags that can be read out with microfluidic barcoding and DNA sequencing. We demonstrate this novel approach by characterizing surface proteins of different cell types at the single-cell level and distinguishing between the cells by their protein expression profiles. DNA-tagged antibodies provide multiple advantages for profiling proteins in single cells, including the ability to amplify low-abundance tags to make them detectable with sequencing, to use molecular indices for quantitative results, and essentially limitless multiplexing. Nature Publishing Group 2017-03-14 /pmc/articles/PMC5349531/ /pubmed/28290550 http://dx.doi.org/10.1038/srep44447 Text en Copyright © 2017, The Author(s) http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Shahi, Payam
Kim, Samuel C.
Haliburton, John R.
Gartner, Zev J.
Abate, Adam R.
Abseq: Ultrahigh-throughput single cell protein profiling with droplet microfluidic barcoding
title Abseq: Ultrahigh-throughput single cell protein profiling with droplet microfluidic barcoding
title_full Abseq: Ultrahigh-throughput single cell protein profiling with droplet microfluidic barcoding
title_fullStr Abseq: Ultrahigh-throughput single cell protein profiling with droplet microfluidic barcoding
title_full_unstemmed Abseq: Ultrahigh-throughput single cell protein profiling with droplet microfluidic barcoding
title_short Abseq: Ultrahigh-throughput single cell protein profiling with droplet microfluidic barcoding
title_sort abseq: ultrahigh-throughput single cell protein profiling with droplet microfluidic barcoding
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5349531/
https://www.ncbi.nlm.nih.gov/pubmed/28290550
http://dx.doi.org/10.1038/srep44447
work_keys_str_mv AT shahipayam absequltrahighthroughputsinglecellproteinprofilingwithdropletmicrofluidicbarcoding
AT kimsamuelc absequltrahighthroughputsinglecellproteinprofilingwithdropletmicrofluidicbarcoding
AT haliburtonjohnr absequltrahighthroughputsinglecellproteinprofilingwithdropletmicrofluidicbarcoding
AT gartnerzevj absequltrahighthroughputsinglecellproteinprofilingwithdropletmicrofluidicbarcoding
AT abateadamr absequltrahighthroughputsinglecellproteinprofilingwithdropletmicrofluidicbarcoding