Nanodroplet Processing Platform for Deep and Quantitative Proteome Profiling of 10-100 Mammalian Cells

Overview

Journal Nat Commun

Specialty Biology

Date 2018 Mar 2

PMID 29491378

Citations 248

Authors

Ying Zhu

Paul D Piehowski

Rui Zhao

Jing Chen

Yufeng Shen

Ronald J Moore

Anil K Shukla

Vladislav A Petyuk

Martha Campbell-Thompson

Clayton E Mathews

Richard D Smith

Wei-Jun Qian

Ryan T Kelly

Affiliations

Soon will be listed here.

Abstract

Nanoscale or single-cell technologies are critical for biomedical applications. However, current mass spectrometry (MS)-based proteomic approaches require samples comprising a minimum of thousands of cells to provide in-depth profiling. Here, we report the development of a nanoPOTS (nanodroplet processing in one pot for trace samples) platform for small cell population proteomics analysis. NanoPOTS enhances the efficiency and recovery of sample processing by downscaling processing volumes to <200 nL to minimize surface losses. When combined with ultrasensitive liquid chromatography-MS, nanoPOTS allows identification of ~1500 to ~3000 proteins from ~10 to ~140 cells, respectively. By incorporating the Match Between Runs algorithm of MaxQuant, >3000 proteins are consistently identified from as few as 10 cells. Furthermore, we demonstrate quantification of ~2400 proteins from single human pancreatic islet thin sections from type 1 diabetic and control donors, illustrating the application of nanoPOTS for spatially resolved proteome measurements from clinical tissues.

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