ArticleAnalytical chemistry2023
Robust and Easy-to-Use One-Pot Workflow for Label-Free Single-Cell Proteomics.
Article in Analytical chemistry, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 69 papers.
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Who cites it
69 citing papers in PubMed.
- High-throughput label-free single-cell proteomics enabled by multicolumn NanoLC.Nature communications · 2026Article
- Pushing the limits of single-cell proteomics to investigate bacterial heterogeneity using bacSCP.Nature communications · 2026Article
- Single-cell proteomic landscape of the developing human brain.Nature biotechnology · 2026Article
- A Parallel Accumulation-Mobility Aligned Fragmentation Strategy Utilizing High-Resolution Ion Mobility for High-Performance Proteomics Analysis.Molecular & cellular proteomics : MCP · 2026Article
- What Are the Practical Applications of Single-Cell Proteomics?Proteomes · 2026Article
- Single-Cell Proteomic Technologies: Tools in the Quest for Principles.Annual review of biophysics · 2026Review
- Low-resolution FAIMS for increased peptide coverage in low-load and single-cell proteomics.Scientific reports · 2026Article
- Single cell proteomic analysis defines discrete neutrophil functional states in human glioblastoma.Nature communications · 2025Article
- Mass spectrometry-based profiling of single-cell histone post-translational modifications to dissect chromatin heterogeneity.Nature communications · 2025Article
- Alternative Ion-Pairing Modifiers Should Be Investigated in Low-Input and Single-Cell Proteomics.Journal of proteome research · 2025Article
- Benchmarking informatics workflows for data-independent acquisition single-cell proteomics.Nature communications · 2025Article
- High-Throughput Label-free Single-Cell Proteomics Enabled by Multicolumn NanoLC with a 5-min Cycle Time.Research square · 2025Article
- Real-Time Eco-AI, Electrophoresis-Correlative Data-Dependent Acquisition with AI-Based Data Processing Broadens Access to Single-Cell Mass Spectrometry Proteomics.Angewandte Chemie (International ed. in English) · 2025Article
- Single-cell proteomics using mass spectrometry.Cell genomics · 2025Review
- Liquid Chromatographic and Mass Spectrometric Methods for Quantitative Proteomic Analysis from Single-Cell and Nanogram-Level Samples.Analytical chemistry · 2025Article
- Sensitive neoantigen discovery by real-time mutanome-guided immunopeptidomics.Nature communications · 2025Article
- Deep Profiling of Oocyte Aging Enabled by Simple One-Step Vial-Based Pretreatment and Single-Cell Proteomics.JACS Au · 2025Article
- Mass Spectrometry-based Solutions for Single-cell Proteomics.Genomics, proteomics & bioinformatics · 2025Review
- Increasing Proteome Coverage Through a Reduction in Analyte Complexity in Single-Cell Equivalent Samples.Journal of proteome research · 2025Article
- Dynamics of Single-Cell Protein Covariation during Epithelial-Mesenchymal Transition.Journal of proteome research · 2025Article
9 more citing papers are in PubMed but not listed here.
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Authors and funding
5 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The analysis of ultralow input samples or even individual cells is essential to answering a multitude of biomedical questions, but current proteomic workflows are limited in their sensitivity and reproducibility. Here, we report a comprehensive workflow that includes improved strategies for all steps, from cell lysis to data analysis. Thanks to convenient-to-handle 1 μL sample volume and standardized 384-well plates, the workflow is easy for even novice users to implement. At the same time, it can be performed semi-automatized using CellenONE, which allows for the highest reproducibility. To achieve high throughput, ultrashort gradient lengths down to 5 min were tested using advanced μ-pillar columns. Data-dependent acquisition (DDA), wide-window acquisition (WWA), data-independent acquisition (DIA), and commonly used advanced data analysis algorithms were benchmarked. Using DDA, 1790 proteins covering a dynamic range of four orders of magnitude were identified in a single cell. Using DIA, proteome coverage increased to more than 2200 proteins identified from single-cell level input in a 20 min active gradient. The workflow enabled differentiation of two cell lines, demonstrating its suitability to cellular heterogeneity determination.
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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.