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High-Throughput Single Cell Proteomics Enabled by Multiplex Isobaric Labeling in a Nanodroplet Sample Preparation Platform

ABSTRACT: Single cell proteomics data from three murine cell lines (Raw, SVEC4-10, C10) and HeLa cell lysate. Samples were digested with trypsin and labeled with TMT 10-Plex on a small scale using nanoPOTS, followed by LC-MS/MS analysis.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Homo Sapiens (ncbitaxon:9606) Mus Musculus (ncbitaxon:10090)

SUBMITTER: Ying Zhu Charles Ansong

PROVIDER: MSV000084110 | MassIVE | Fri Jul 19 12:06:00 BST 2019

SECONDARY ACCESSION(S): PXD014684

REPOSITORIES: MassIVE

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High-Throughput Single Cell Proteomics Enabled by Multiplex Isobaric Labeling in a Nanodroplet Sample Preparation Platform.

Dou Maowei M Clair Geremy G Tsai Chia-Feng CF Xu Kerui K Chrisler William B WB Sontag Ryan L RL Zhao Rui R Moore Ronald J RJ Liu Tao T Pasa-Tolic Ljiljana L Smith Richard D RD Shi Tujin T Adkins Joshua N JN Qian Wei-Jun WJ Kelly Ryan T RT Ansong Charles C Zhu Ying Y

Analytical chemistry 20190925 20

Effective 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 ...[more]

PMID: 31509397

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