ArticleMolecular & cellular proteomics : MCP2016
Label-free Quantification of Proteins in Single Embryonic Cells with Neural Fate in the Cleavage-Stage Frog (Xenopus laevis) Embryo using Capillary Electrophoresis Electrospray Ionization High-Resolution Mass Spectrometry (CE-ESI-HRMS).
Article in Molecular & cellular proteomics : MCP, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 50 papers.
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Who cites it
50 citing papers in PubMed, 89 citations in OpenAlex.
- Single-cell proteomic landscape of the developing human brain.Nature biotechnology · 2026Article
- Electrospray Regime Controls Sensitivity and Quantitative Performance in Micro- and Nano-Flow CE-ESI-MS.Analytical chemistry · 2026Article
- Subcellular mass spectrometry reveals proteome remodeling in an asymmetrically dividing (frog) embryonic stem cell.Proceedings of the National Academy of Sciences of the United States of America · 2026Article
- 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
- Online Coupling of Acoustic Droplet Levitation with Capillary Electrophoresis Mass Spectrometry Enables Midair Enrichment for Improved Proteomic Sensitivity.Analytical chemistry · 2025Article
- A Multipoint Validation of Quantification in Capillary Electrophoresis Mass Spectrometry Proteomics: Isobaric Multiplexing with Tandem Mass Tags.Analytical chemistry · 2025Article
- Data-Independent Acquisition Shortens the Analytical Window of Single-Cell Proteomics to Fifteen Minutes in Capillary Electrophoresis Mass Spectrometry.Journal of proteome research · 2025Article
- Development and Validation of RoboCap, a Robotic Capillary Platform to Automate Capillary Electrophoresis Mass Spectrometry En Route to High-Throughput Single-Cell Proteomics.Analytical chemistry · 2024Article
- Coupling Microdroplet-Based Sample Preparation, Multiplexed Isobaric Labeling, and Nanoflow Peptide Fractionation for Deep Proteome Profiling of the Tissue Microenvironment.Analytical chemistry · 2024Article
- Microanalytical Mass Spectrometry with Super-Resolution Microscopy Reveals a Proteome Transition During Development of the Brain's Circadian Pacemaker.Analytical chemistry · 2023Article
- Monitoring and modelling the dynamics of the cellular glycolysis pathway: A review and future perspectives.Molecular metabolism · 2022Review
- Separation methods in single-cell proteomics: RPLC or CE?International journal of mass spectrometry · 2022Article
- Extracellular vesicles expressing CEACAM proteins in the urine of bladder cancer patients.Cancer science · 2022Article
- Profiling 26,000 Aplysia californica neurons by single cell mass spectrometry reveals neuronal populations with distinct neuropeptide profiles.The Journal of biological chemistry · 2022Article
- Cell-Lineage Guided Mass Spectrometry Proteomics in the Developing (Frog) Embryo.Journal of visualized experiments : JoVE · 2022Article
- Applications of Tandem Mass Spectrometry (MS/MS) in Protein Analysis for Biomedical Research.Molecules (Basel, Switzerland) · 2022Review
- Single-Cell Mass Spectrometry of Metabolites and Proteins for Systems and Functional Biology.Neuromethods · 2022Article
- Capillary Electrophoresis Mass Spectrometry for Scalable Single-Cell Proteomics.Frontiers in chemistry · 2022Article
- Proteomics for Low Cell Numbers: How to Optimize the Sample Preparation Workflow for Mass Spectrometry Analysis.Journal of proteome research · 2021Article
- In Vivo Subcellular Mass Spectrometry Enables Proteo-Metabolomic Single-Cell Systems Biology in a Chordate Embryo Developing to a Normally Behaving Tadpole (X. laevis)*.Angewandte Chemie (International ed. in English) · 2021Article
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Quantification of protein expression in single cells promises to advance a systems-level understanding of normal development. Using a bottom-up proteomic workflow and multiplexing quantification by tandem mass tags, we recently demonstrated relative quantification between single embryonic cells (blastomeres) in the frog (Xenopus laevis) embryo. In this study, we minimize derivatization steps to enhance analytical sensitivity and use label-free quantification (LFQ) for single Xenopus cells. The technology builds on a custom-designed capillary electrophoresis microflow-electrospray ionization high-resolution mass spectrometry platform and LFQ by MaxLFQ (MaxQuant). By judiciously tailoring performance to peptide separation, ionization, and data-dependent acquisition, we demonstrate an ∼75-amol (∼11 nm) lower limit of detection and quantification for proteins in complex cell digests. The platform enabled the identification of 438 nonredundant protein groups by measuring 16 ng of protein digest, or <0.2% of the total protein contained in a blastomere in the 16-cell embryo. LFQ intensity was validated as a quantitative proxy for protein abundance. Correlation analysis was performed to compare protein quantities between the embryo and n = 3 different single D11 blastomeres, which are fated to develop into the nervous system. A total of 335 nonredundant protein groups were quantified in union between the single D11 cells spanning a 4 log-order concentration range. LFQ and correlation analysis detected expected proteomic differences between the whole embryo and blastomeres, and also found translational differences between individual D11 cells. LFQ on single cells raises exciting possibilities to study gene expression in other cells and models to help better understand cell processes on a systems biology level.
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Registered trials
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