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[Preprint]. 2023 May 31:2023.01.06.522908. [Version 4] doi: 10.1101/2023.01.06.522908

High Throughput Single Cell Proteomic Analysis of Organ Derived Heterogeneous Cell Populations by Nanoflow Dual Trap Single Column Liquid Chromatography

Simion Kreimer, Aleksandra Binek, Blandine Chazarin, Jae Hyung Cho, Ali Haghani, Alexandre Hutton, Eduardo Marbán, Mitra Mastali, Jesse G Meyer, Thassio Mesquita, Yang Song, Jennifer Van Eyk, Sarah Parker
PMCID: PMC9881989  PMID: 36711540

Abstract

Identification and proteomic characterization of rare cell types within complex organ derived cell mixtures is best accomplished by label-free quantitative mass spectrometry. High throughput is required to rapidly survey hundreds to thousands of individual cells to adequately represent rare populations. Here we present parallelized nanoflow dual-trap single-column liquid chromatography (nanoDTSC) operating at 15 minutes of total run time per cell with peptides quantified over 11.5 minutes using standard commercial components, thus offering an accessible and efficient LC solution to analyze 96 single-cells per day. At this throughput, nanoDTSC quantified over 1,000 proteins in individual cardiomyocytes and heterogenous populations of single cells from aorta.

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