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Instruments present in our facility are:
SELDI-TOF-MS (surface enhanced laser
desorption/ionisation-time of flight-mass spectrometer)
This method uses proteinchip® arrays (Biorad) with different selective surfaces such as cation or anion exchange surfaces, hydrophobic surfaces and metal
binding surfaces. Also antibodies, specific proteins and DNA can be bound
to chips to study protein-protein and protein-DNA interactions. Cell lysate,
plasma or urine is applied on the selective surface and, after washing,
a subset of proteins is specifically bound. The chip is analyzed in a
TOF-MS
which generates a protein
spectrum of the different molecular masses present on the proteinchip.
This technology is suitable for research into molecular mechanisms of
disease and biomarker identification.
2D GEL and DIGE
Fluorescence 2-D difference gel electrophoresis (DIGE)
uses spectrally resolvable dyes to label protein samples prior to 2-D
electrophoresis. By using different fluorescent dyes to separately label
protein samples multiple samples can be co-separated and visualized on a
single 2-D gel. This fluorescent multiplexing approach is compatible with
mass spectrometry and overcomes many of the disadvantages of traditional
2-D analyses. A broad dynamic range provides more accurate quantitative
data than traditional 2-D silver staining techniques while rapid image
overlay simplifies image analysis and improves comparative accuracy.

Biacore 3000
This instrument is based on the
SPR
(Surface Plasmon Resonance) technology that enables real-time detection
and monitoring of biomolecular binding events (e.g. protein-protein interactions).
Biological processes are, for a large part, determined by protein-protein
interactions which play a role in the pathogenesis of acquired or genetic
diseases. Furthermore, protein-protein interactions, e.g. hormone-receptor
interactions, are key targets for drug design.
Surface Plasmon Resonance (SPR) technology: Upon binding of proteins
a change in refractive index is measured which represents the change in
molecular mass.
Multidimensional HPLC-MS
High-performance-liquid chromatography (HPLC) coupled to electrospray
ionisation mass spectrometry (ESI-MS) is a powerful technique to separate
peptides and proteins of lower molecular weight (appr. < 20 kDa). This
technology will be further developed for large-scale analysis of biological
fluids for new biomarkers of disease and other applications requiring
high sample throughput.
Applied
Biosystems Voyager DE Pro mass spectrometer with MALDI source
Matrix Assisted Laser Desorption
Ionization (MALDI) allows a quick and simple determination of the molecular
weight of a protein, a peptide, and even mixtures of proteins and peptides.
Combined with enzymatic digestion it will deliver a peptide mass fingerprint
of a protein, wich can be used for searching a data base and identification
of a protein. MALDI makes use of a matrix which absorbs the light of the
laser puls that is fired on the sample. Sample ions are then liberated from
the mixture of matrix and sample, and ions are detected by means of a time
of flight mass spectrometer. The Voyager offers high sensitivity and high
mass accuracy, and is simple to use in the work flow of a proteomics
facility.

ABI-MDS/Sciex
QSTAR hybrid mass spectrometer with ion sources for electrospray, nanospray,
MALDI and SELDI
The QSTAR is a versatile
high-performance research mass spectrometer which comprises a quadrupole
section for mass separation, a collision cell for ion fragmentation, and a
Time-of -flight section for accurate mass determination. It can be combined
with a nanobore HPLC system intended for the separation of peptide mixtures
at high sensitivity. When the MALDI or SELDI sources are used, the ease of
use of these ionization methods is combined with the extensive series of
options for ion fragmentation and accurate mass measurement, aimed at the
determination of the amino acid sequence of peptides and the structure
elucidation of new biomarkers.

ChipLC-Q-TOF
MS system
The Agilent 6510 Q-TOF LC/MS delivers a ten-fold
sensitivity advantage, allowing the detection of critical, low-level
components. At the same time, fast spectral acquisition allows the
acquisition of higher-quality data and provides more information from
complex samples. The system is equipped with the
Agilent 1200 Series HPLC-Chip/MS
system. This is a new microfluidic chip-based technology for nanospray
LC/MS. Based on the HPLC-Chip and HPLC-Chip Cube MS interface, it provides a
new level of nanospray MS sensitivity, robustness, ease of use and
reliability.


HPLC-Chip Cube MS interface
LTQ
Orbitrap high resolution MS
The LTQ Orbitrap (Thermo Electron) can
deliver low-ppm mass accuracy
and extremely high resolution, all within a time scale compatible with
nano-LC separations. Therefore, the Orbitrap is a useful instrument for many
proteomics applications that involve protein identification of low abundant
proteins with high accuracy.

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