Abseq: Ultrahigh-throughput single cell protein profiling with droplet microfluidic barcoding
Year: 2017
Type: article
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.
Source: Scientific Reports
DOI: 10.1038/srep44447
Institutions: QB3, University of California, San Francisco
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $2,690
Funders: National Science Foundation, National Institutes of Health, Defense Advanced Research Projects Agency, Advanced Research Projects Agency, National Human Genome Research Institute, National Institute of General Medical Sciences, National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institute of Biomedical Imaging and Bioengineering
Awards: Molecular Biophysics Training Grant, G1863003419, Gigapixel digital PCR in Giant Unilamellar Vesicles, G3637141366, G382641700, Accurate and unbiased single cell sequencing with digital droplet MDA, G5206177008, CAREER: Massively-parallel single-cell genomics and transcriptomics with droplet microfluidics, G549111027, Microfluidic immunoprofiling for biomarker discovery in rheumatoid arthritis, G6671297155, Accurate, long-read sequencing with droplet-microfluidic barcoding, Integrative Program in Complex Biological System (ipCBS), G9000068819
OpenAlex ID: W2603592147
