Proteomic Analysis of Single Mammalian Cells Enabled by Microfluidic Nanodroplet Sample Preparation and Ultrasensitive NanoLC‐MS
Ying Zhu(Pacific Northwest National Laboratory), Gérémy Clair(Pacific Northwest National Laboratory), William Chrisler(Pacific Northwest National Laboratory), Yufeng Shen(Pacific Northwest National Laboratory), Rui Zhao(Pacific Northwest National Laboratory), Anil Shukla(Pacific Northwest National Laboratory), Ronald Moore(Pacific Northwest National Laboratory), Ravi Misra(University of Rochester Medical Center), Gloria Pryhuber(University of Rochester Medical Center), Richard Smith(Pacific Northwest National Laboratory), Charles Ansong(Pacific Northwest National Laboratory), Ryan Kelly(Pacific Northwest National Laboratory)
Cited by 269Open Access
Abstract
We report on the quantitative proteomic analysis of single mammalian cells. Fluorescence-activated cell sorting was employed to deposit cells into a newly developed nanodroplet sample processing chip, after which samples were analyzed by ultrasensitive nanoLC-MS. An average of circa 670 protein groups were confidently identified from single HeLa cells, which is a far greater level of proteome coverage for single cells than has been previously reported. We demonstrate that the single-cell proteomics platform can be used to differentiate cell types from enzyme-dissociated human lung primary cells and identify specific protein markers for epithelial and mesenchymal cells.
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