Efficient and Sensitive Sample Preparation, Separations, and Data Acquisition for Label-Free Single-Cell Proteomics.

Thy Truong, Ximena Sanchez-Avila, Kei G I Webber, S Madisyn Johnston, Ryan T Kelly

Abstract read

In one paragraph

Article in Methods in molecular biology (Clifton, N.J.), 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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0citing papers in PubMed

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1 · What the graph read from it

What it found

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2 · The registry

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0 citing papers in PubMed.

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4 · The record

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5 · Who and what money

Authors and funding

5 authors.

Thy TruongDepartment of Chemistry and Biochemistry, Brigham Young University, Provo, UT, USA.

Ximena Sanchez-AvilaDepartment of Chemistry and Biochemistry, Brigham Young University, Provo, UT, USA.

Kei G I WebberDepartment of Chemistry and Biochemistry, Brigham Young University, Provo, UT, USA.

S Madisyn JohnstonDepartment of Chemistry and Biochemistry, Brigham Young University, Provo, UT, USA.

Ryan T KellyDepartment of Chemistry and Biochemistry, Brigham Young University, Provo, UT, USA. ryan.kelly@byu.edu.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

We describe a sensitive and efficient workflow for label-free single-cell proteomics that spans sample preparation, liquid chromatography separations, and mass spectrometry data acquisition. The Tecan Uno Single Cell Dispenser provides rapid cell isolation and nanoliter-volume reagent dispensing within 384-well PCR plates. A newly developed sample processing workflow achieves cell lysis, protein denaturation, and digestion in 1 h with a single reagent dispensing step. Low-flow liquid chromatography coupled with wide-window data-dependent acquisition results in the quantification of nearly 3000 proteins per cell using an Orbitrap Exploris 480 mass spectrometer. This approach greatly broadens accessibility to sensitive single-cell proteome profiling for nonspecialist laboratories.

Indexed as

ProteomicsSingle-Cell AnalysisChromatography, LiquidHumansMass SpectrometryProteinsProteomeProteinsProteomeHP D100 single cell dispenserLabel-free quantificationNanoLCOne-step workflowSingle-cell proteomicsWide window acquisition

Identifiers

PMID38907148

DOI10.1021/acs.jproteome.0c00629

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.