RRC ID 69161
著者 Gebreyesus ST, Siyal AA, Kitata RB, Chen ES, Enkhbayar B, Angata T, Lin KI, Chen YJ, Tu HL.
タイトル Streamlined single-cell proteomics by an integrated microfluidic chip and data-independent acquisition mass spectrometry.
ジャーナル Nat Commun
Abstract Single-cell proteomics can reveal cellular phenotypic heterogeneity and cell-specific functional networks underlying biological processes. Here, we present a streamlined workflow combining microfluidic chips for all-in-one proteomic sample preparation and data-independent acquisition (DIA) mass spectrometry (MS) for proteomic analysis down to the single-cell level. The proteomics chips enable multiplexed and automated cell isolation/counting/imaging and sample processing in a single device. Combining chip-based sample handling with DIA-MS using project-specific mass spectral libraries, we profile on average ~1,500 protein groups across 20 single mammalian cells. Applying the chip-DIA workflow to profile the proteomes of adherent and non-adherent malignant cells, we cover a dynamic range of 5 orders of magnitude with good reproducibility and <16% missing values between runs. Taken together, the chip-DIA workflow offers all-in-one cell characterization, analytical sensitivity and robustness, and the option to add additional functionalities in the future, thus providing a basis for advanced single-cell proteomics applications.
巻・号 13(1)
ページ 37
公開日 2022-1-10
DOI 10.1038/s41467-021-27778-4
PII 10.1038/s41467-021-27778-4
PMID 35013269
PMC PMC8748772
MeSH Animals Cell Line, Tumor Cell Separation Humans Lab-On-A-Chip Devices* Lung Neoplasms Mass Spectrometry / methods* Microfluidics / methods* Proteome Proteomics / methods* Reproducibility of Results Workflow
IF 12.121
リソース情報
ヒト・動物細胞 PC-9(RCB4455)