Copper sulfate (CuSO4.5H2O), d -( +)-glucose, and polyvinylpyrrolidone (PVP) were obtained from Cicarelli (Buenos Aires, Argentina). Sodium hydroxide was provided by Anedra (Buenos Aires, Argentina). MALDI matrix, 2,5-dihydroxybenzoic acid (DHB), was purchased from Bruker Daltonics Gmbh (Bremen, Germany). Immunoglobulin G from human serum (IgG, 56,834-25MG), albumin from bovine serum (BSA, A3059), cytochrome C from equine heart (Cyt C), acetonitrile (ACN), and trifluoroacetic acid (TFA) were purchased from Sigma-Aldrich (Stockholm, Sweden). All reagents were of analytical grade and used without further purification. Ultrapure water with a resistivity of 18.2 MΩ cm at 25 °C (Millipore Synergy® 185 system, Bedford, MA, USA) was used throughout the experiments. For analysis of glycopeptides, the IgG standard was subjected to trypsin digestion and the digested mixture had a final concentration of 1 mg mL−1 [18 (link)].
Maldi matrix
Manufactured by Bruker
Sourced in Germany
The MALDI matrix is a solid chemical compound used in matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. It facilitates the ionization of analyte molecules, allowing their detection and analysis by the mass spectrometer.
Automatically generated - may contain errors
Lab products found in correlation
Trifluoroacetic acid tfa, by Merck Group (2 mentions)
Flexcontrol v3, by Bruker (2 mentions)
Formic acid, by Merck Group (2 mentions)
Maldi biotyper 3, by Bruker (2 mentions)
Acetonitrile acn, by Merck Group (1 mentions)
Synergy 185 system, by Merck Group (1 mentions)
Maldi tof ms, by Bruker (1 mentions)
Ammonium phosphate, by Merck Group (1 mentions)
Flexanalysis 3, by Bruker (1 mentions)
Mtp 384 polished steel target plate, by Bruker (1 mentions)
α cyano 4 hydroxycinnamic acid, by Bruker (1 mentions)
Peptide calibration standard 2, by Bruker (1 mentions)
Maldi tof mass spectrometer, by Bruker (1 mentions)
Acetonitrile, by Carl Roth (1 mentions)
4 protocols using maldi matrix
1
Glycopeptide Analysis Protocol for IgG
2
MALDI-TOF MS Microbial Identification
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One loop of colony, or pellet, obtained at the end of the extraction procedure was spread using a pipette tip onto the MALDI-TOF MS (Bruker Daltonik GmbH) steel plate spots and allowed to air dry. Next, one microlitre of 70% formic acid (Sigma-Aldrich, Darmstadt, Germany) was added and allowed to dry. Last, the spots were covered with the MALDI matrix (10 mg/mL α-cyano-4-hydroxy-cinnamic acid in 50% acetonitrile/0.1% trifluoroacetic acid; Bruker Daltonik GmbH). All samples were analysed in duplicate. Mass spectra were acquired in a MALDI Microflex LT/SH bench-top mass spectrometer (Bruker Daltonik GmbH) equipped with a 60 Hz nitrogen laser. FlexControl v.3.0 software (Bruker Daltonik GmbH) was used to acquire the spectra, and the MALDI Biotyper 3.1 (Bruker Daltonik GmbH) was used for real-time interpretation and identification of the microorganisms. According to the manufacturer’s instructions (Bruker Daltonik GmbH), a score of more than 2.0 indicates good species identification, a score of between 1.7 and 2.0 indicates good genus identification, and a score of <1.7 indicates unreliable identification [12 (link)].
3
MALDI-TOF MS Protein Identification Protocol
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The pellet obtained at the end of the extraction procedure was spread with a pipette tip on the MALDI-TOF MS steel plate spots and allowed to dry. One microlitre of 70% formic acid (Sigma-Aldrich, USA) was added to the sample and allowed to air-dry. The spots were then covered with the MALDI matrix (10 mg/mL acyano-4-hydroxy-cinnamic acid in 50% acetonitrile/0.1% trifluoroacetic acid; Bruker Daltonik GmbH). Samples were analysed in duplicate. Spectra were acquired in a MALDI Microflex LT/SH bench-top mass spectrometer (Bruker Daltonik GmbH) equipped with a 60 Hz nitrogen laser. FlexControl v.3.0 software (Bruker Daltonik GmbH) was used to acquire the spectra and the MALDI Biotyper 3.1 (Bruker Daltonik GmbH) for real-time interpretation and identification of the microorganisms. According to the manufacturer, a score >2.0 indicates species identification, a score between 1.7 and 2.0 indicates genus identification, and a score <1.7 indicates unreliable identification.
4
MALDI-TOF Analysis of Endopeptidase Digests
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MS-based Endopep analysis was done in MS-positive ion reflector mode utilizing an autoflex speed matrix-assisted laser desorption/ionisation time-of-flight (MALDI-TOF) mass spectrometer (Bruker Daltonics, Bremen, Germany) equipped with a smartbeam laser. Sample (2 µL) was mixed with 18 µL of MALDI-Matrix (5 mg/mL of α-Cyano-4hydroxycinnamic acid, Bruker Daltonics) in 0.1% trifluoroacetic acid (TFA, Sigma-Aldrich, Seelze, Germany) and 50% acetonitrile (Carl Roth, Karlsruhe, Germany) in HPLC water, 1 mM ammoniumphosphate (Merck, Darmstadt, Germany). Of this mixture 1 µL was spotted onto a MTP 384 polished steel target plate (Bruker Daltonics). For matrix suppression, deflection was set to m/z 500, mass spectra were acquired over the mass range m/z 600 to 4800. External mass calibration was performed with peptide calibration standard II (Bruker Daltonics). Each spectrum represents an average of 4,000 laser shots. Spectra were processed by flexAnalysis 3.4 software (Bruker Daltonics).
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