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Download fileHigh-Throughput Single Cell Proteomics Enabled by Multiplex Isobaric Labeling in a Nanodroplet Sample Preparation Platform
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posted on 2019-09-24, 20:13 authored by Maowei Dou, Geremy Clair, Chia-Feng Tsai, Kerui Xu, William B. Chrisler, Ryan L. Sontag, Rui Zhao, Ronald J. Moore, Tao Liu, Ljiljana Pasa-Tolic, Richard D. Smith, Tujin Shi, Joshua N. Adkins, Wei-Jun Qian, Ryan T. Kelly, Charles Ansong, Ying ZhuEffective
extension of mass spectrometry-based proteomics to single
cells remains challenging. Herein we combined microfluidic nanodroplet
technology with tandem mass tag (TMT) isobaric labeling to significantly
improve analysis throughput and proteome coverage for single mammalian
cells. Isobaric labeling facilitated multiplex analysis of single
cell-sized protein quantities to a depth of ∼1 600 proteins
with a median CV of 10.9% and correlation coefficient of 0.98. To
demonstrate in-depth high throughput single cell analysis, the platform
was applied to measure protein expression in 72 single cells from
three murine cell populations (epithelial, immune, and endothelial
cells) in <2 days instrument time with over 2 300 proteins
identified. Principal component analysis grouped the single cells
into three distinct populations based on protein expression with each
population characterized by well-known cell-type specific markers.
Our platform enables high throughput and unbiased characterization
of single cell heterogeneity at the proteome level.
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TMTanalysis throughput2 300 proteinsNanodroplet Sample Preparation Platform2 300Principal component analysiscell heterogeneitycorrelation coefficientHigh-Throughput Single Cell Proteomics EnabledMultiplex Isobaricmultiplex analysistandem mass tagcell analysiscell-sized protein quantitiesCVproteome levelmicrofluidic nanodroplet technologymass spectrometry-based proteomicsprotein expressionmurine cell populationsmeasure protein expressionproteome coverage