MassIVE MSV000084612

Complete Public PXD016410

Ultrasensitive Top-Down Proteomics Analysis of Low Number of Mammalian Cells Using a Nanodroplet Sample Processing Platform

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Description

We developed an ultrasensitive top down platform by incorporating a microfluidic sample preparation system, termed nanoPOTS (Nanodroplet Processing in One pot for Trace Samples), into a top down workflow. We determined that a unique combination of a nonionic detergent DDM with urea as a protein extraction buffer can significantly improve both protein extraction efficiency and sample recovery. The nanoPOTS-based top down platform can reproducibly and quantitatively identify ~170 to ~620 proteoforms from ~70 to ~770 HeLa cells containing < ~100 ng total proteins. Data was searched with TopPIC and MSPathFinder. [doi:10.25345/C5HM3H] [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: top-down ; proteomics ; microfluidics ; nanodroplet

Contact

Principal Investigators:
(in alphabetical order) 		Ying Zhu, Pacific Northwest National Laboratory, USA
Submitting User: alchemistmatt

Publications

Zhou M, Uwugiaren N, Williams SM, Moore RJ, Zhao R, Goodlett D, Dapic I, Paša-Toli? L, Zhu Y.
Sensitive Top-Down Proteomics Analysis of a Low Number of Mammalian Cells Using a Nanodroplet Sample Processing Platform.
Anal. Chem. Epub 2020 May 6.

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