Proteomic Analysis of Single Mammalian Cells Enabled by Microfluidic Nanodroplet Sample Preparation and Ultrasensitive NanoLC‐MS

• Published in: Angewandte Chemie (International ed.) Vol. 57; no. 38; pp. 12370 - 12374
• Main Authors: Zhu, Ying, Clair, Geremy, Chrisler, William B., Shen, Yufeng, Zhao, Rui, Shukla, Anil K., Moore, Ronald J., Misra, Ravi S., Pryhuber, Gloria S., Smith, Richard D., Ansong, Charles, Kelly, Ryan T.
• Format: Journal Article
• Language: English
• Published: Germany Wiley Subscription Services, Inc 17.09.2018; Wiley
• Edition: International ed. in English
• Subjects: cell typing; droplets; Fluorescence; Lungs; Mammalian cells; Mammals; Mesenchyme; Microfluidics; Proteins; Proteomics; Sample preparation; single-cell proteomics; ultrasensitive LC-MS; ultrasensitive LC-MS; cell typing; single-cell proteomics; droplets; microfluidics
• ISSN: 1433-7851; 1521-3773
• DOI: 10.1002/anie.201802843

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. Single‐cell proteomics: A microfluidic platform coupled to nanoLC‐MS was developed to enable quantitative proteomic analysis of single mammalian cells containing only 0.1–0.2 ng of total protein. Label‐free cell differentiation was enabled by quantifying protein expression in individual cells.