Development of an Automated, Ultra-Rapid Bottom-Up Proteomics Workflow Utilizing Alginate-Based Hydrogels Article Swipe
YOU?
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· 2024
· Open Access
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· DOI: https://doi.org/10.1021/acs.analchem.4c04846
A new approach to sample preparation and enzymatic digestion in bottom-up proteomics has been developed using alginate-based hydrogel entrapment of enzymes. This hydrogel facilitates rapid and room-temperature digestions with multienzyme capabilities. Three methodologies were tested: within microcentrifuge tubes, in situ pipette tips, and automated robotic liquid handling. Factorial experimental design identified a 1 h, room temperature, pepsin-trypsin dual-enzyme digestion as optimal for sequence coverage and protein group identification, comparable to a gold-standard overnight proteomic protocol. This method promises significant advancements in proteomic analysis by enhancing reusability, speed, throughput, convenience, and cost-effectiveness, without hindering digestion efficiency.
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- Type
- article
- Language
- en
- Landing Page
- https://doi.org/10.1021/acs.analchem.4c04846
- https://pubs.acs.org/doi/pdf/10.1021/acs.analchem.4c04846?ref=article_openPDF
- OA Status
- hybrid
- Cited By
- 4
- References
- 42
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W4404235245