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Issue 3, 2021
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Ultrasensitive single-cell proteomics workflow identifies >1000 protein groups per mammalian cell

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Abstract

We report on the combination of nanodroplet sample preparation, ultra-low-flow nanoLC, high-field asymmetric ion mobility spectrometry (FAIMS), and the latest-generation Orbitrap Eclipse Tribrid mass spectrometer for greatly improved single-cell proteome profiling. FAIMS effectively filtered out singly charged ions for more effective MS analysis of multiply charged peptides, resulting in an average of 1056 protein groups identified from single HeLa cells without MS1-level feature matching. This is 2.3 times more identifications than without FAIMS and a far greater level of proteome coverage for single mammalian cells than has been previously reported for a label-free study. Differential analysis of single microdissected motor neurons and interneurons from human spinal tissue indicated a similar level of proteome coverage, and the two subpopulations of cells were readily differentiated based on single-cell label-free quantification.

Graphical abstract: Ultrasensitive single-cell proteomics workflow identifies >1000 protein groups per mammalian cell

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Supplementary files

Article information


Submitted
01 Jul 2020
Accepted
15 Nov 2020
First published
17 Nov 2020

This article is Open Access
All publication charges for this article have been paid for by the Royal Society of Chemistry

Chem. Sci., 2021,12, 1001-1006
Article type
Edge Article

Ultrasensitive single-cell proteomics workflow identifies >1000 protein groups per mammalian cell

Y. Cong, K. Motamedchaboki, S. A. Misal, Y. Liang, A. J. Guise, T. Truong, R. Huguet, E. D. Plowey, Y. Zhu, D. Lopez-Ferrer and R. T. Kelly, Chem. Sci., 2021, 12, 1001 DOI: 10.1039/D0SC03636F

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