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A Plasma Sample Preparation for Mass Spectrometry using an Automated Workstation
Published on: April 24, 2020
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A Plasma Sample Preparation for Mass Spectrometry using an Automated Workstation.
Qin Fu1, Casey W Johnson1, Bhagya K Wijayawardena2
1Advanced Clinical Biosystems Institute, Smidt Heart institute, Cedars Sinai Medical Center.
Journal of Visualized Experiments : Jove
|May 12, 2020
Summary
This study introduces an improved automated workflow for proteomics sample preparation, reducing steps and enhancing control for more reliable mass spectrometry data. The optimized process increases efficiency and precision in protein and peptide quantification.
Area of Science:
- Proteomics
- Analytical Chemistry
- Biochemistry
Background:
- Proteomics sample preparation for mass spectrometry involves multiple liquid transfer and incubation steps.
- Current methods can be time-consuming and prone to variability, impacting data reliability.
- Automation offers a potential solution to increase efficiency and reproducibility.
Purpose of the Study:
- To develop a more efficient and controlled automated workflow for enzymatic protein digestion in proteomics.
- To reduce the number of liquid transfer steps and pipetting time.
- To improve the overall reproducibility and precision of mass spectrometry-based quantification.
Main Methods:
- Implemented a redesigned automated workflow reducing liquid transfers from nine to six by reagent combination.
- Utilized selective tip pipetting with a 96-channel pipetting head to minimize pipetting time.
- Integrated a fully enclosed system for enhanced temperature and environmental control.
- Incorporated stable isotope labeled peptides and β-galactosidase for process monitoring and quality control.
Main Results:
- Reduced liquid transfers and pipetting time, increasing potential throughput with 45 deck positions.
- Achieved improved intra-assay and inter-assay precision (CV < 20%) for LC-MS quantification.
- Completed the digestion of 96 samples in approximately 5 hours.
- Demonstrated enhanced reproducibility and reduced potential for contamination.
Conclusions:
- The enhanced automated workflow significantly improves efficiency and control in proteomics sample preparation.
- This optimized process leads to more reliable and reproducible LC-MS based protein and peptide quantification.
- The system offers a robust solution for high-throughput, precise proteomic analyses.
