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Enhanced protein identification using graphite-modified MALDI plates for offline LC-MALDI-MS/MS bottom-up proteomics
Anthony Maus1, Rudolph Mignon1, Franco Basile1
1Department of Chemistry, University of Wyoming, 1000 E. University Ave., Laramie, WY 82072, United States.
Analytical Biochemistry
|January 13, 2018
Summary
Coating matrix-assisted laser desorption/ionization (MALDI) plates with graphite powder significantly enhances protein identification in bottom-up proteomics. This cost-effective method boosts peptide and protein detection rates in liquid chromatography-MALDI tandem mass spectrometry (LC-MALDI-MS/MS) workflows.
Area of Science:
- Proteomics
- Analytical Chemistry
- Biochemistry
Background:
- Offline liquid chromatography-matrix assisted laser desorption/ionization tandem mass spectrometry (LC-MALDI-MS/MS) is a cost-effective bottom-up proteomics technique.
- Improving protein identification is crucial for comprehensive proteomic analysis.
Purpose of the Study:
- To enhance protein identification capabilities in bottom-up proteomics using LC-MALDI-MS/MS.
- To develop a simple, low-cost method to increase peptide signal intensity and protein inference.
Main Methods:
- Developed a method involving coating MALDI plates with graphite powder.
- Applied the modified plates in an offline LC-MALDI-MS/MS bottom-up proteomic workflow.
- Utilized the Escherichia coli proteome for validation.
Main Results:
- Graphite-modified MALDI plates increased peptide identifications by 50-135%.
- Concomitant increase of 21%-105% in inferred protein identifications was observed.
- Identified factors contributing to improved peptide and protein identifications.
Conclusions:
- Graphite coating of MALDI plates is an effective strategy to enhance protein identification in LC-MALDI-MS/MS.
- The method is low-cost, easy to implement, and compatible with existing workflows without adding analysis time.
- This approach significantly improves the efficiency of bottom-up proteomics.
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