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Whole-body Mass Spectrometry Imaging by Infrared Matrix-assisted Laser Desorption Electrospray Ionization (IR-MALDESI)
Published on: March 24, 2016
A new, modular mass calibrant for high-mass MALDI-MS
Simon Weidmann1, Konstantin Barylyuk, Nadezhda Nespovitaya
1Department of Chemistry and Applied Biosciences, ETH Zurich, Zurich, Switzerland.
Analytical Chemistry
|February 12, 2013
Summary
Researchers developed new protein calibrants for high-mass analysis using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). These recombinantly produced maltodextrin-binding protein (MBP) standards cover a wide mass range, improving protein mass spectrometry accuracy.
Area of Science:
- Proteomics
- Analytical Chemistry
- Biochemistry
Background:
- Accurate mass calibration is crucial for high-mass protein analysis via matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS).
- Existing calibration standards often lack suitability for the high m/z ratios encountered in high-mass protein studies.
Purpose of the Study:
- To develop and evaluate novel, well-suited calibration standards for high-mass protein analysis using MALDI-MS.
- To establish a set of calibrants covering the m/z range of 40-400 kDa.
Main Methods:
- Recombinant expression of concatenated polyproteins based on maltodextrin-binding protein (MBP).
- Introduction of specific recognition sites for selective enzymatic cleavage.
- Generation of MBP oligomers (MBP2, MBP3, MBP4, MBP6) as calibration standards.
- MALDI-MS analysis of generated protein calibrants.
Main Results:
- Concatenated MBP polyproteins demonstrated suitability as calibration standards for high-mass MALDI-MS.
- A set of calibrants covering the m/z range of 40-400 kDa was successfully generated.
- Despite differing only in mass, equimolar mixtures of these calibrants did not produce equal signal intensities on a high-mass detector.
Conclusions:
- Recombinantly produced MBP-based polyproteins are effective calibrants for high-mass MALDI-MS.
- Further investigation is needed to understand and address signal intensity variations in equimolar mixtures of these calibrants.
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