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Differential Scanning Calorimetry (µ-DSC) Theory

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µ-DSC is a well-stablished method for thermodynamic analysis of biomolecules, due to combined high data reproducibility and method sensitivity, and the “structural fingerprint” character of thermograms that is unique for each biomolecule. In addition, µ-DSC provides a direct and label-free measurement of thermal stability without the same solvents restrictions that apply to spectroscopic methods.

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μ-DSC is highly effective to demonstrate biosimilarity, which is done by the direct comparison between the thermogram (a structural “fingerprint”) of the reference product and a biosimilar candidate. In addition, it provides high data reproducibility and sensitivity to factors influencing thermal stability and conformational changes.

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Temperature stability is a critical parameter in drug product development, especially during formulation development or within biosimilarity studies. μ-DSC is the gold standard analytical method for thermal stability analysis of biomolecules. Additionally, ZentriForce Pharma offers static and dynamic light scattering (SLS and DLS) methods that, when applied during a temperature ramp, are fast formulation screening tools for thermal stability characterization.

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ZentriForce Pharma was founded as a service provider specialized in hydrodynamic techniques used for biopharmaceutical development. The focus lies on gene therapy products and biosimilarity studies. For further information follow the link to our company page.

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ZentriForce Pharma is your expert in analytical services, with focus on hydrodynamic techniques. Please follow the link to find out about our analytical services. 

ZentriForce Pharma offers a wide range of analytical techniques. Please follow the link to our analytical portfolio. 

ZentriForce Pharma’ strength is not restricted to expertise in analytical services. Our scientist are also actively involved in the further advances in the theory of hydrodynamic techniques. Please follow the link to a detailed description of the theory behind our analytical services, including some of our own research.

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Microcal Differential Scanning Calorimetry (µ-DSC)

Differential scanning calorimetry (DSC) induces thermal transitions of biomolecules and registers their heat capacity change (∆Cp) as a function of temperature. As a result, the melting temperatures (Tm) and the transition enthalpies (∆H) of thermal events are determined.

DSC is a reliable and accurate technique for thermal stability analysis of biomolecules that undergo one or more temperature – dependent transitions. ZentriForce is equipped with a Microcal DSC (µ-DSC) instrument for analysis of proteinaceous samples and liposomal formulations.

μ-DSC thermogram of a protein
Depiction of a μ-DSC thermogram of a protein unfolding process.

µ-DSC is ideally suited for solutions that interfere with optical analysis methods including turbid, colored or particulate samples. It stands out as a validation tool for techniques such as CD and FT-IR spectroscopy.

Additionally, DSC is a direct measurement; it requires no labels. Further, it measures the unfolding process directly, while many other techniques merely detect the unfolded versus the folded state.

Description of µ-DSC Experiments

In a DSC measurement, two cells are heated simultaneously but independently from each other at a specific heating rate. One cell holds the sample, while the other cell (reference) holds the solvent. This heating process induces conformational or phase transitions in the sample.

The transitions are accompanied by a change in heat capacity of the molecules in the sample. If the reaction is endotherm, the sample requires more heat than the reference because noncovalent inter- or intramolecular bonds must be broken by heat influx (the heat capacity of the molecule increases). Therefore, more energy is transferred to the sample cell than to the reference cell to keep both at the same temperature.

As a result, DSC provides the sample’s heat capacity as a function of temperature and thus establishes the thermodynamic profile of this conformational change.

This  thermodynamic profile is used as a higher order structure (HOS) comparability study of biopharmaceuticals, for instance, in biosimilar development, to show that a biosimilar’s thermodynamic profile is highly similar to that of the reference product.

Further µ-DSC key applications include:

  • Stability screenings for formulation development
  • Batch-to-batch comparisons
  • Membrane and lipids characterization

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