Activities, equipment and facilities

The Macromolecular Interaction Analysis Unit is available for scientific collaborations or for fee-for-service by the university community and by biotechnological or pharmaceutical companies.

    SPR analyses of interactions with measurements of kinetic parameters: association and dissociation rate (Kon and Koff), kinetic and steady state dissociation constant (Kd).
    SPR analyses of drug/target interactions: direct interaction with measurement of kinetic parameters or competition experiments with calculation of the inhibitory potency (ID50) of the drug on the target ligand/receptors interaction.
    Characterization of the biological consequences of the interactions analyzed:in vitro cell culture assays, ex vivo assays for endothelial physio-pathology, oncology and cardiovascular diseases.

At the Macromolecular Interaction Analysis Unit are operative one BIACORE X-100 apparatus and one BIACORE X apparatus fully equipped with dedicated software for macromolecular interaction analysis.
BIACORE X-100

    Sample handling: automatic, maximum 15 samples, 24 h unattended operation
    Molecular Weight Detection:  Down to 100
    Refractive Index range:  1.33-1.40
    Sample Volume: 20 to 30 µl
    Sample Type: LMW drug candidates to high molecular weight proteins (also DNA, RNA, polysaccharides, lipids, cells and viruses)
    Analysis time per sample: 5-15 min
    Detection limit:0.1 nM for  >10.000 Da analysis nM for > 10.000 Da analysis
    Kinetics range:
    Association range constant: 103-107 M-1s-1BIACORE X-100
    Dissociation range constant: 10-5-0.1 s-1
    Flow rate range: 1-100 μl/min
    Analysis temperature: 4 - 40°C
    Reference subtraction:automatic
    Number of flow Cells: 2
    Baseline Drift:<0.3 RU/min
    Baseline Noise: <0.1 RU

BIACORE X

    Sample handling: Manual sample loading
    Molecular weight detection: >180 D
    Refractive index range: 1.33 - 1.40
    Sample volume:+20 μl
    Sample Type: LMW drug candidates to high molecular weight proteins (also DNA, RNA, polysaccharides, lipids, cells and viruses)
    Detection limit:10 pM (myoglobin, 17.000 Da, sandwich assay
    Kinetics range:
    Association range constant: 103-5x106 M-1s-1
    Dissociation range constant: 10-5-0.1 s-1BIACORE X
    Flow rate range: 1 - 100 μl/min
    Analysis temperature: 4 - 40°C
    In-line reference subtraction: yes
    Number of flow cells: 2
    Baseline Drift:<0.3 RU/min
    Baseline Noise: <0.6 RU

Also, the Macromolecular Interaction Analysis Unit has fulfilled the requirements for microorganism/gene manipulation, bacteria growth, protein purification and analysis and cellular biology studies: protein and RNA electrophoresis apparatus, PCR apparatus, ultrafiltration apparatus, FPLC system, cold/sterile rooms, biohazard/chemical hoods, CO2 incubators for mammalian cell cultures, stereomicroscope and inverted fluorescence microscope for time-lapse studies.

From molecular interactions to biological consequences