One-Tip enables comprehensive proteome coverage in minimal cells and single zygotes
Zilu Ye1,2, Pierre Sabatier3,4, Javier Martin-Gonzalez5
1State Key Laboratory of Common Mechanism Research for Major Diseases, Suzhou Institute of Systems Medicine, Chinese Academy of Medical Sciences & Peking Union Medical College, Suzhou, China. yzl@ism.pumc.edu.cn.
Nature Communications
|March 20, 2024
Summary
One-Tip is a new proteomics method simplifying sample preparation for reproducible protein identification. This efficient workflow enhances proteomic analysis across diverse sample types, from cells to extracellular vesicles.
Area of Science:
- Proteomics
- Mass Spectrometry
- Biochemistry
Background:
- Mass spectrometry (MS)-based proteomics workflows are complex, multi-step processes.
- These workflows face challenges including sample loss, reproducibility issues, high costs, and required expertise.
Purpose of the Study:
- To introduce One-Tip, a novel proteomics methodology.
- To simplify sample preparation and enhance data-independent acquisition (DIA) analysis.
- To demonstrate the broad applicability of One-Tip across various sample types.
Main Methods:
- Integration of one-pot sample preparation with narrow-window data-independent acquisition (nDIA).
- Utilized the Uno Single Cell Dispenser™ for single-cell proteomics.
- Applied the workflow to HeLa cells, single mouse embryo cells, and extracellular vesicles (EVs) from blood plasma.
Main Results:
- Reproducible identification of >9000 proteins from ~1000 HeLa cells.
- Identification of ~6000 proteins in single cells from early mouse embryos.
- Identification of >3000 proteins per HeLa cell using single-cell proteomics.
- Identification of >3000 proteins from 16 ng of EVs from blood plasma.
Conclusions:
- One-Tip significantly simplifies proteomics sample processing.
- The methodology enables high-depth and reproducible protein identification across diverse biological samples.
- One-Tip expands the capabilities of proteomics in terms of depth, throughput, and sample versatility.
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