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Characterization Quality Control Pharmacokinetics

Biomarker Discovery MALDI-8020

Profiling Cancer Cells Using a benefits

Benchtop MALDI-TOF MS System Cell Line Optimization
click here • Enables rapid and highly sensitive benchtop profiling.
• Easy maintenance and low running costs
Operating Principle and Features extracellular vesicles from regular lymph node cells metastasized from
colon cancer versus from lymph node cells with elevated chemotherapy • Can search a wide range of molecular weights for nucleic acids, proteins, and molecules.
Compared to quadrupole or magnetic sector mass spectrometers, resistance.
matrix-assisted laser desorption/ionization time-of-flight (MALDI- Protein was collected from the extracellular vesicles obtained from
TOF) mass spectrometers offer the advantage of both a broad mass cultivated cells and then the MALDI-8020 system was used to obtain a Culture
measurement range and fast measurement speed. They are especially mass spectrum. Alpha-cyano-4-hydroxycinnamic acid (CHCA) was used
well-suited for measuring nucleic acids, proteins, and molecules. as a matrix. eMSTAT Solution was used to analyze the resulting mass
Though a benchtop system with a compact size, the MALDI-8020 spectrum by multivariate analysis (Fig. 1).
offers world-class resolution and sensitivity levels. In addition, a shorter
vacuum evacuation time is achieved by increasing the laser speed and Results
modifying the exhaust system, and a significantly shorter measurement
time is achieved by increasing stage speed. Components derived from the protein were detected in the m/z 2,000
to 25,000 range of the extracellular vesicle mass spectrum. Multivariate
Measurement Method analysis score plot results discriminated between groups with resistance
versus groups with sensitivity to fluorouracil. From the peak matrix, Purification
Cancer cells can metastasize throughout the body by means of peaks that characterized chemotherapy resistance were detected in the
extracellular vesicles. Therefore, a MALDI-TOF MS system was used m/z 2,000 to 7,000 range (Fig. 2).
to profile differences in the expression level of proteins derived from
Specifications

Instrument MALDI-8020
Cell culture
Mass range m/z 1 to 500,000
Collect supernatant
Mass resolution >5,000 FWHM
Spin at Spin at Spin at
500 g 16,500 g 120,000 g Characterization
Sensitivity >250 amol
cells cell debris Extracellular
vesicles
Mass accuracy <20 ppm with internal calibration, <150 ppm with external calibration
Sequential Ultracentrifugation Spot sample and matrix Acceleration voltage 15 kV
Sample preparation MALDI-8020 eMSTAT Solution
Laser Solid-state laser wavelength: 355 nm Repetition frequency: 50, 100, or 200 Hz (variable)
Fig. 1 Workflow for Extracellular Vesicle Analysis
Flight distance 850 mm
Detector Electron Multiplier
Mass spectrum Ion source cleaning Includes automatic cleaning functionality (depending on built-in solid-state laser)
Peak Matrix： Sample plate Disposable FlexiMass-DS and stainless steel FlexiMass-SR Quality Control
This shows the group-speci c
peaks detected.
Red : Peaks detected in Operating noise <55 dB
all mass spectra
Pink : Peaks detected in Main unit power supply Single-phase 100 V AC, 50/60 Hz, 1 kVA
only some mass spectra
Green : Peaks not detected in Dimensions W 600 mm × D 745 mm × H 1,055 mm (excluding protrusions)
any mass spectra
Score Plot Weight 86 kg
● Primary colon cancer
● Resistant to 5 mol/L uorouracil Operating environment Temperature: 18 to 28 °C Humidity: Max. 70 % (with no condensation)
● Resistant to 25 mol/L uorouracil
● Resistant to 125 mol/L uorouracil Data analysis software eMSTAT Solution
Fig. 2 PLS-DA Analysis Results for Four Extracellular Vesicle Groups Data analysis Univariate analysis t-Test, Mann-Whitney U-Test, ANOVA (analysis of variance) Pharmacokinetics
functionality
Conclusion Application Examples (Shimadzu Application News No.) Multivariate analysis PCA (principal component analysis), PLS-DA
Discriminant analysis Support Vector Machine (SVM), Random Forest
Using the MALDI-8020 system in combination with statistical analysis • Profiling protein expression in tissue
software shows its potential for use in biomarker discovery research. • Analyzing glycans and glycopeptides (B113) Other Dynamic grouping
This type of protein profiling method can be expected to be useful for • Analyzing the primary structures of protein (B105) Display functionality Multivariate analysis Peak Matrix, Box Plot, ROC, AUC, Score/Loading Plot, Dendrogram
less invasive cancer diagnosis or for monitoring chemotherapy.
Discriminant analysis Discriminant analysis results (Group, Score) superimpose points for unknown samples on a score plot Others
Input/output data Input Peak list (ASCII, JCAMP, or mzML format)
The extracellular vesicles and cell extracts from primary colon cancer and metastasized lymph node subclones with resistance to 5, 25, and 125 μM concentrations of 5-fluorouracil (FU) were provided by Output Peak list (txt format), data analysis results (xlsm format), graph screenshot
Dr. Gerald Stubiger of the Medical University of Vienna.

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