Biophysical Studies

Analytical Ultracentrifuge (AUC)

Instrument: Optima AUC from Beckman Coulter Life Sciences with absorbance and interference detection modules, two rotors, An-60 Ti and An-50 Ti are available.

Applications: Analytical ultracentrifugation is a highly versatile analytical method and brings valuable benefits during the research and development of (bio)pharmaceutical drug products.

When (macro)molecules in solution are subject to a centrifugal force, they begin to settle at a certain velocity. With a mathematical correlation between sedimentation behavior and hydrodynamic properties, scientists can determine many critical parameters of a drug substance or drug product for a variety of biopharmaceutical applications from analyzing small peptides to investigating macromolecular interactions.

Biolayer interferometry (OCTET)

Instrument: Forte Bio OCTET Red 96e with temperature control

Applications: Analysis of biomolecular kinetics (rates of association and dissociation), binding affinities, and quantification of macromolecules.

Information provided

Circular Dichroism Spectrometer (CD)

Instrument: Chirascan CD spectrometer from Applied Photophysics.

Applications: Analysis of secondary structure of proteins and chiral molecules, binding and melting studies.

Measuring CD over the near-UV range (>250 nm) generates a CD fingerprint of the tertiary structure of biomolecules. The fingerprint is influenced by the nature of the surrounding environment of aromatic side chains of the amino-acids tryptophan, phenylalanine and tyrosine. The presence of disulfide bonds may also influence the final fingerprint.

Differential Scanning Calorimetry (DSC)

Instrument: MicroCal PEAQ-DSC

Applications: Thermal stability of proteins and other biomolecules.

DSC is a powerful tool for polymer analysis, and generates data that provides insight into:

Curing Reactions, Chemical Reactions, & Curie Point (Curie Temperature)
Crystallization Structure & Fusion
OIT Oxidative Induction Times
Thermal & Oxidative Stability
Decomposition Temperature
Component Identification
Melting Point & Range
Glass Transition
Sample Purity
Heat Capacity

Dynamic Light Scattering (DLS)

Instrument: DynaPro Nanostar from Wyatt Technology

Applications: Analysis of homogeneity of biological samples (protein solutions etc), determination of hydrodynamic radius, molecular masses, and oligomeric states.

DSC is a powerful tool for polymer analysis, and generates data that provides insight into:

Accurate, reliable and repeatable particle size analysis in one or two minutes.
Multi-Angle Dynamic Light Scattering (MADLS®) improves the resolution of DLS and provides angular independent size results.
Simple, material independent measurement of per peak particle concentration.
Mean size only requires knowledge of the viscosity of the liquid.
Simple or no sample preparation, high concentration, turbid samples can be measured directly using NIBS.
Size measurement of sizes from <1nm to >10um
Size measurement of molecules from MW < 1000Da.
Low volume requirement (as little as 3µL).

Isothermal Titration Calorimetry (ITC)

Instrument: MicroCal PEAQ-ITC.

Applications: Measurement of binding affinities, enthalpy, and entropy changes.

ITC has advantages over other techniques such as fluorescence assays, NMR and SPR for studying complex formation in terms of ease of use and cost. It does not require any fluorescent probes or radioactive tags for data analysis. Immobilization and chemical modification of protein is not required. Also, it does not have limitations associated with clarity of the solution, molecular weight, temperature or pH. It is one of the best methods for determining the thermodynamic parameters of ligand binding.

Microscale Thermophoresis (MST)

Instrument: Monolith NT.115 (Blue/Red) from NanoTemper.

Applications: Measurement of binding affinities

The technique is based on the motion of molecules along local temperature gradients, measured by fluorescence changes. A spatial temperature difference (ΔT) leads to a mass flow of molecules in the temperature-elevated region. The Soret coefficient, also known as thermophoresis, was defined by ST: chot = ccold * exp(− ST ΔT ) and quantified the relation of temperature gradient and the molecule flow. The thermophoresis of a protein typically differs significantly from the thermophoresis of a protein-ligand complex due to binding-induced changes in size, charge, and hydration shell.

Multimode Plate Reader

Instrument: Spectramax iD5 from Molecular Devices.

Applications: The SpectraMax® iD5 Multi-Mode Microplate Readers measure absorbance, fluorescence, and luminescence.

In addition, the iD5 reader measures TRF and FP and can be expanded to include TR-FRET, HTRF®, BRET, dual luciferase reporter assays with injectors, and western blot detection.

Stopped Flow

Instrument: SX20 Stopped-Flow spectrometer from Applied Photophysics.

Applications: This technique involves two reactants held in separate reservoirs that are prevented from freely flowing by syringe pumps. The reaction is initiated by depressing the reactant syringes, and thus releasing the reactants into the connecting “mixing chamber” where the solutions are mixed. The reaction is monitored by observing the change in absorbance of the reaction solution as a function of time.

As the reaction progresses it fills the “stop syringe” which then expands until it hits a block at the point when the reaction has reached a continuous flow rate, thereby stopping the flow and the reaction, and thus allowing the researcher to calculate the exact initial rate of reaction.

Hydrogen Deuterium Exchange LC-MS

H/D exchange gives information about the solvent accessibility of various parts of the molecule, and thus the tertiary structure of the protein. Detection can be measured by NMR, mass spectrometry.

Analytical Ultracentrifuge (AUC)

Biolayer interferometry (OCTET)

Instrument: Forte Bio OCTET Red 96e with temperature control

Applications: Analysis of biomolecular kinetics (rates of association and dissociation), binding affinities, and quantification of macromolecules.

Information provided

Circular Dichroism Spectrometer (CD)

Instrument: Chirascan CD spectrometer from Applied Photophysics.

Applications: Analysis of secondary structure of proteins and chiral molecules, binding and melting studies.

Differential Scanning Calorimetry (DSC)

Instrument: MicroCal PEAQ-DSC

Applications: Thermal stability of proteins and other biomolecules.

DSC is a powerful tool for polymer analysis, and generates data that provides insight into:

Curing Reactions, Chemical Reactions, & Curie Point (Curie Temperature)
Crystallization Structure & Fusion
OIT Oxidative Induction Times
Thermal & Oxidative Stability
Decomposition Temperature
Component Identification
Melting Point & Range
Glass Transition
Sample Purity
Heat Capacity

Dynamic Light Scattering (DLS)

Instrument: DynaPro Nanostar from Wyatt Technology

Applications: Analysis of homogeneity of biological samples (protein solutions etc), determination of hydrodynamic radius, molecular masses, and oligomeric states.

DSC is a powerful tool for polymer analysis, and generates data that provides insight into:

Accurate, reliable and repeatable particle size analysis in one or two minutes.
Multi-Angle Dynamic Light Scattering (MADLS®) improves the resolution of DLS and provides angular independent size results.
Simple, material independent measurement of per peak particle concentration.
Mean size only requires knowledge of the viscosity of the liquid.
Simple or no sample preparation, high concentration, turbid samples can be measured directly using NIBS.
Size measurement of sizes from <1nm to >10um
Size measurement of molecules from MW < 1000Da.
Low volume requirement (as little as 3µL).

Isothermal Titration Calorimetry (ITC)

Instrument: MicroCal PEAQ-ITC.

Applications: Measurement of binding affinities, enthalpy, and entropy changes.

Microscale Thermophoresis (MST)

Instrument: Monolith NT.115 (Blue/Red) from NanoTemper.

Applications: Measurement of binding affinities

Multimode Plate Reader

Instrument: Spectramax iD5 from Molecular Devices.

Applications: The SpectraMax® iD5 Multi-Mode Microplate Readers measure absorbance, fluorescence, and luminescence.

In addition, the iD5 reader measures TRF and FP and can be expanded to include TR-FRET, HTRF®, BRET, dual luciferase reporter assays with injectors, and western blot detection.

Stopped Flow

Instrument: SX20 Stopped-Flow spectrometer from Applied Photophysics.

Biophysical Studies

Analytical Ultracentrifuge (AUC)

Biolayer interferometry (OCTET)

Circular Dichroism Spectrometer (CD)

Differential Scanning Calorimetry (DSC)

Dynamic Light Scattering (DLS)

Fluorimeter

Isothermal Titration Calorimetry (ITC)

Microscale Thermophoresis (MST)

Multimode Plate Reader

Stopped Flow

Hydrogen Deuterium Exchange LC-MS

Analytical Ultracentrifuge (AUC)

Biolayer interferometry (OCTET)

Circular Dichroism Spectrometer (CD)

Differential Scanning Calorimetry (DSC)

Dynamic Light Scattering (DLS)

Fluorimeter

Isothermal Titration Calorimetry (ITC)

Microscale Thermophoresis (MST)

Multimode Plate Reader

Stopped Flow

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