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Sci Rep. 2017 Mar 14;7:44447. doi: 10.1038/srep44447.

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

Author information

1
Department of Bioengineering and Therapeutic Sciences and California Institute for Quantitative Biosciences (QB3), University of California, San Francisco, San Francisco, CA 94158, USA.
2
Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94158, USA.

Abstract

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.

PMID:
28290550
PMCID:
PMC5349531
DOI:
10.1038/srep44447
[Indexed for MEDLINE]
Free PMC Article

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