ArticleChemical science2018
Nanowell-mediated two-dimensional liquid chromatography enables deep proteome profiling of <1000 mammalian cells.
Article in Chemical science, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed, 41 citations in OpenAlex.
- The pursuit of ultrasensitive phosphoproteomics to unravel signalling in rare cells.Communications biology · 2025Review
- Exploring new frontiers in type 1 diabetes through advanced mass-spectrometry-based molecular measurements.Trends in molecular medicine · 2024Review
- Parallel sample processing for mass spectrometry-based single cell proteomics.Analytica chimica acta · 2024Article
- Coupling Microdroplet-Based Sample Preparation, Multiplexed Isobaric Labeling, and Nanoflow Peptide Fractionation for Deep Proteome Profiling of the Tissue Microenvironment.Analytical chemistry · 2024Article
- Repurposed 3D Printer Allows Economical and Programmable Fraction Collection for Proteomics of Nanogram Scale Samples.Analytical chemistry · 2024Article
- Efficient and Sensitive Sample Preparation, Separations, and Data Acquisition for Label-Free Single-Cell Proteomics.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Bottom-Up Proteomics: Advancements in Sample Preparation.International journal of molecular sciences · 2023Review
- In-Depth Mass Spectrometry-Based Proteomics of Formalin-Fixed, Paraffin-Embedded Tissues with a Spatial Resolution of 50-200 μm.Journal of proteome research · 2022Article
- Seamlessly Integrated Miniaturized Filter-Aided Sample Preparation Method to Fractionation Techniques for Fast, Loss-Less, and In-Depth Proteomics Analysis of 1 μg of Cell Lysates at Low Cost.Analytical chemistry · 2022Article
- High-Density Nanowells Formation in Ultrafast Laser-Irradiated Thin Film Metallic Glass.Nano-micro letters · 2022Article
- Review
- MicroPOTS Analysis of Barrett's Esophageal Cell Line Models Identifies Proteomic Changes after Physiologic and Radiation Stress.Journal of proteome research · 2021Article
- Proteomic/transcriptomic analysis of erythropoiesis.Current opinion in hematology · 2021Review
- Ultrasensitive single-cell proteomics workflow identifies >1000 protein groups per mammalian cell.Chemical science · 2020Article
- Single-cell Proteomics: Progress and Prospects.Molecular & cellular proteomics : MCP · 2020Review
- Molecular Dynamics Simulation-assisted Ionic Liquid Screening for Deep Coverage Proteome Analysis.Molecular & cellular proteomics : MCP · 2020Article
- Accurate Identification of Deamidation and Citrullination from Global Shotgun Proteomics Data Using a Dual-Search Delta Score Strategy.Journal of proteome research · 2020Article
- Review of Three-Dimensional Liquid Chromatography Platforms for Bottom-Up Proteomics.International journal of molecular sciences · 2020Review
- Improved Single-Cell Proteome Coverage Using Narrow-Bore Packed NanoLC Columns and Ultrasensitive Mass Spectrometry.Analytical chemistry · 2020Article
- High-Throughput Single Cell Proteomics Enabled by Multiplex Isobaric Labeling in a Nanodroplet Sample Preparation Platform.Analytical chemistry · 2019Article
Corrections and comments
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Authors and funding
13 authors at 2 institutions in 2 countries.
Funding
Abstract
Multidimensional peptide separations can greatly increase the depth of coverage in proteome profiling. However, a major challenge for multidimensional separations is the requirement of large biological samples, often containing milligram amounts of protein. We have developed nanowell-mediated two-dimensional (2D) reversed-phase nanoflow liquid chromatography (LC) separations for in-depth proteome profiling of low-nanogram samples. Peptides are first separated using high-pH LC and the effluent is concatenated into 4 or 12 nanowells. The contents of each nanowell are reconstituted in LC buffer and collected for subsequent separation and analysis by low-pH nanoLC-MS/MS. The nanowell platform minimizes peptide losses to surfaces in offline 2D LC fractionation, enabling >5800 proteins to be confidently identified from just 50 ng of HeLa digest. Furthermore, in combination with a recently developed nanowell-based sample preparation workflow, we demonstrated deep proteome profiling of >6000 protein groups from small populations of cells, including ∼650 HeLa cells and 10 single human pancreatic islet thin sections (∼1000 cells) from a pre-symptomatic type 1 diabetic donor.
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Registered trials
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.