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On-Chip Sample Preparation Using a ChipFilter Coupled to NanoLC-MS/MS for Bottom-Up Proteomics
Massamba M Ndiaye1, Ha Phuong Ta1, Giovanni Chiappetta1
1Biological Mass Spectrometry and Proteomics (SMBP), ESPCI Paris, Université PSL, CNRS, 75005 Paris, France.
Journal of Proteome Research
|April 29, 2020
Summary
A new microfluidic device, ChipFilter Proteolysis (CFP), miniaturizes protein sample preparation and digestion for bottom-up proteomics. This innovative reactor enhances proteome coverage using less material and in less time compared to commercial methods.
Area of Science:
- Proteomics
- Analytical Chemistry
- Microfluidics
Background:
- Bottom-up proteomics relies heavily on efficient sample preparation.
- Existing microfluidic devices often struggle with complex samples and integrated processing/digestion.
- Miniaturization offers potential for improved analytical performance in proteomics.
Purpose of the Study:
- To introduce the ChipFilter Proteolysis (CFP) microfluidic device for miniaturized protein sample processing and digestion.
- To demonstrate the CFP device's capability in handling complex proteomic samples.
- To compare the CFP device's performance against commercial ultracentrifugation cartridges.
Main Methods:
- Design and fabrication of a two-chamber microfluidic device with an integrated filtration membrane.
- Utilized the CFP device for cell lysis, protein concentration, and rapid chemical/enzymatic digestion.
- Analyzed complex samples including yeast protein extract and whole human cell proteome.
Main Results:
- The CFP device successfully processed complex proteomic samples.
- Achieved superior proteome coverage compared to commercial ultracentrifugation cartridges.
- Required 10 times less starting material and was 8 times faster than the commercial method.
Conclusions:
- The CFP microfluidic device is an effective reactor for miniaturized proteomics sample preparation and digestion.
- This approach offers significant advantages in terms of efficiency, speed, and material usage.
- The CFP device represents a promising advancement for bottom-up proteomics analysis.
