View All Molecular Biophysics at Instruct
The platform provides instruments and services leading to the characterization of biomolecular interactions (determination of affinity, kinetic and thermodynamic parameters) using isothermal titration calorimetry (ITC), surface plasmon resonance (SPR), microscale thermophoresis (MST) and analytical ultracetrifugation (AUC).
Analytical ultracentrifugation (AUC) is a powerful technique for the characterisation of biomolecules and their interactions in a solution. It is applicable to proteins, nucleic acids, lipids, polysaccharides, viruses and nanoparticles. Analytical ultracentrifugation is used to determine the degree of the sample heterogeneity, the oligomeric state (or molar mass, respectively), the shape of the particle and to characterize biomolecular interactions (stoichiometry, Kd). Instrument: ProteomeLab XL-I (Beckman Coulter). More information on the instrument and the sample requirements can be found here.
Isothermal titration calorimetry (ITC) is used to measure the binding affinity and thermodynamics of a wide variety of biomolecular interactions (those including e.g. proteins, nucleic acids, lipids or small ligands), and to study the enzyme kinetics. ITC measures the heat released or absorbed during the binding event and a complete thermodynamic profile of the interaction can be obtained in a single experiment (stoichiometry, Kd, ∆H and ∆S). ITC requires no labeling, accommodates wide range of affinities and the measurement can be performed over a range of conditions (temperature, ionic strength, pH, etc.). Instrument: The platform offers both the manual (VP-ITC, Malvern) and the automated, high-throughput calorimeter (PEAQ-ITC Automated, Malvern). More information on the instrument and the sample requirements can be found here.
Microscale thermophoresis (MST) is based on measuring changes of the mobility of molecules in the microscopic temperature gradients. It allows for determination of affinity and stoichiometry of the biomolecular interactions. The interaction is measured through the changes in hydration shell, charge or size of the fluorescently-labeled molecule. MST can be used for measuring interactions between molecules of different sizes and masses (ions to viruses) with a very low sample consumption. Instruments: Monolith NT.115 (Nanotemper), Monolith NT.115 Pico (Nanotemper) More information on the instrument and the sample requirements can be found here.
The SPR biosensor system exploits the phenomenon of surface plasmon resonance to monitor the interactions between molecules in a real time. One of the binding partners is immobilized on the sensor chip surface, while the other is passed over the surface in a solution. SPR method is used to determine the specificity, affinity and kinetics of the interaction. The method finds its application in different fields of science, e.g. in structural biology or biotherapeutic and drug discovery research. Instrument: Biacore T200 (GE Healthcare) More information on the instrument and the sample requirements can be found here.
Analytical ultracentrifugation (Beckman Coulter ProteomeLab XL-I)
Analytical ultracentrifugation (AUC) is a powerful technique for the characterization of biomolecules and their interactions in a solution. It is applicable to proteins, nucleic acids, lipids, polysaccharides, viruses and nanoparticles. Analytical ultracentrifugation has a broad applicability in life sciences, including biomolecular interactions, and allows for determination of Kd and stoichiometry of both self-associating (protein oligomerization) and hetero-associating systems (e.g. protein-protein, protein-DNA interactions).
More information on the instrument and the sample requirements can be found here.
Isothermal titration calorimetry (Malvern Auto-iTC200 and VP-ITC)
Isothermal titration calorimetry (ITC) is used to measure the binding affinity and thermodynamics of a wide variety of biomolecular interactions (those including e.g. proteins, nucleic acids, lipids or small ligands), and to study the enzyme kinetics. ITC measures the heat released or absorbed during the binding event and a complete thermodynamic profile of the interaction can be obtained in a single experiment (stoichiometry, Kd, ∆H and ∆S). ITC requires no labeling, accommodates wide range of affinities and the measurement can be performed over a range of conditions (temperature, ionic strength, pH, etc.).
The platform offers both the manual and the automated, high-throughput calorimeter. More information on the instruments and the sample requirements can be found here: Auto-iTC200 VP-ITC
Microscale thermophoresis (Nanotemper Monolith NT.115)
Microscale thermophoresis (MST) is based on measuring changes of the mobility of molecules in the microscopic temperature gradients. It allows for determination of affinity and stoichiometry of the biomolecular interactions. The interaction is measured through the changes in hydration shell, charge or size of the fluorescently-labeled molecule. MST can be used for measuring interactions between molecules of different sizes and masses (ions to viruses) with a very low sample consumption.
More information on the instrument and the sample requirements can be found here.
Surface plasmon resonance (Biacore T200)
The SPR biosensor system exploits the phenomenon of surface plasmon resonance to monitor the interactions between molecules in a real time. One of the binding partners is immobilized on the sensor chip surface, while the other is passed over the surface in a solution. SPR method is used to determine the specificity, affinity and kinetics of the interaction. The method finds its application different fields of science, e.g. in structural biology or biotherapeutic and drug discovery research.
More information on the instrument and the sample requirements can be found here.