1. Investice do rozvoje vzdělávání
Overview of the use of MALDI-TOF MS and MALDI MSI within the NANOTEAM project
Roman Guráň
Investice do rozvoje vzdělávání

2. Bruker New ultrafleXtremeTM
The innovative smartbeam-II™ laser enables ultra-high data acquisition speed in both MS and MS/MS at full systems performance. The well-established proprietary smartbeam laser provides unprecedented analytical and matrix flexibility in workflows from protein tissue imaging, intact proteins analysis, Glycoproteomics, biologics or oligo QC, to LC-MALDI proteomics - all fully enabled at Hz repetition rates.
The new ultrafleXtreme now enables laser focus diameters down to 10 µm for high spatial resolution imaging without pixel overlap. Importantly, outstanding spectral quality and signal intensity are maintained at even the smallest laser beam diameters.
The novel FlashDetector™ combined with a minimum of 4 GHz digitizer and latest advances in electronics provide unmatched mass resolving power up to 40,000 and 1 ppm mass accuracy for highest confidence.
The novel and unique laser-irradiation self-cleaning Ion Source ensures robust, long-term highest-performance operation. Very long MALDI laser lifetime in combination with automated source cleaning in just minutes leads to high uptime and low maintenance costs¨.
Latest TOF/TOF technology: The high efficiency and sensitivity of the LID-LIFT process delivers MS/MS spectra with nominal mass resolution for peptides. Typically, full MS/MS data sets can be acquired with up to 1000 Hz laser repetition rate from low fmol levels within seconds. The Bruker bioinformatics software is perfectly adapted to analyze and visualize the match between the raw spectra and annotated peptide and protein structures.

3. Applications in our lab
Identification of proteins, bacteria and other
MALDI mass spectrometry imaging of different tissues

4. Results under CEITEC project
Influence of microbiome species representation in the hard-to-heal wounds on disease severity and treatment duration
MALDI-TOF MS was used for identification of bacteria and then the dendrograms were created for comparing identified bacteria.

5. Results under CEITEC project
Structural effects and nanoparticle size are essential for quantum dots-metallothionein complex formation
MALDI-TOF MS was used for characterization of metallothionein.
Peak No.
Molecular mass found (Da)
Molecular mass expected (Da)
Identification 1
6111
6113
Apo MT-2C 2
6131 - 3
6139
N-acetylated and C-methylated apo MT-2A 4
6171
6169
N-acetylated and C-methylated apo MT-2C 5
6210
6209
Na+ ion of C-methylated apo MT-2D 6
6231
6230
N-acetylated and C-methylated apo MT-2D 7
6321
6322
adduct of Na+ ion of N-acetylated apo MT-2B with molecule DHB (matrix) 8
6362
6363
adduct of Na+ ion of C-methylated apo MT-2D with molecule DHB (matrix) 9
6418
6417
adduct of Na+ ion of N-acetylated apo MT-2E with molecule DHB (matrix) adduct of N-acetylated and C-methylated apo MT-2D with molecule DHB (matrix) B
Intensity (a.u.)
6 000
6 200
6 400
6 600
2 000
4 000
8 000
5 800 1 5 4 6 7 23 8 9
m/z

6. Results under CEITEC project
Differences in urinary proteins related to surgical margin status after radical prostatectomy
MALDI-TOF MS was used for identification of urinary proteins by peptide mass fingerprinting (PMF) after 2D gel electrophoresis.

7. Results under SIX project
Study of Linkage between Glutathione Pathway and the Antibiotic Resistance of Escherichia coli from patients’ swabs
MALDI-TOF MS was used for comparison of different E.coli samples and for detection of glutathione (GSH).

8. Results under SIX project
Growth properties and ability to form biofilms by Staphylococcus aureus and MRSA after the treatment of antibiotics with SeNPs
MALDI-TOF MS was used for comparing protein profiles of S. aureus and MRSA after application of antibiotics with SeNPs.

9. Results under SIX project
The substitution of organic matrices by carbon nanomaterials for MALDI-TOF MS analyses of low molecular weight analytes like glutathione
MALDI-TOF MS was used for testing carbon nanomaterials as new matrices for low molecular weight analytes.

10. Future work
MALDI MSI of different tumors – we will focus especially on detection of metallothioneins
Identification of other bacterial samples from Trauma hospital of Brno

11. Investice do rozvoje vzdělávání
Acknowledgements
Investice do rozvoje vzdělávání 11

12. Thank you for your attention
Investice do rozvoje vzdělávání