Biophysical Characterization of Macromolecules


Various analytical techniques are used to study and analyze biopharmaceutical molecules. Biophysical characterization is critical to understanding details of the structure - function relationship of biological macromolecules.

Microcalorimetry techniques: Isothermal titration calorimetry (ITC) and differential scanning calorimetry (DSC), are powerful tools for in-depth characterization of molecular binding events and structural stability. DSC and ITC generate comprehensive thermodynamic profiles for protein domain structures and the energetics of inter- and intra-molecular binding events. These techniques are a direct approach for determining stability and affinity through thermodynamics in a single experiment.

Native Assays:  Domain structures and stability, binding constants, stoichiometry and enthalpy of reactions can be generated through microcalorimetry without any molecular modifications such as labeling or immobilization to facilitate assay development. 

Applications for DSC and ITC data include:

  • Structural domain determination
  • Biopharmaceutical pre-formulation stability studies
  • Secondary screening of protein-protein interactions
  • Excipient evaluation in formulations
  • Specific/non-specific binding determinations


Featured Application Notes:

Localized Conformation Analysis of Mutated Human IgG1 by HDX MS and nanoDSC - This technology brief describes the process of probing mAb conformation change caused by amino acid mutation using both Hydrogen Deuterium Exchange (HDX MS, or HXMS) and nano Differential Scanning Calorimetry (DSC) technologies.

Determining Thermal Stability of Antibodies with a Nano DSC - Antibodies or immunoglobulins are a specific type of glycoprotein. The Nano DSC is an essential tool for scientists working with antibodies to characterize the structure-function relationships that may exist.

Characterizing Protein-Protein Interactions by ITC - This note examines the application of isothermal titration calorimetry for quantitatively measuring the thermodynamic properties driving protein-protein interactions.

Featured Webinars:

A Practical Approach to Microcalorimetry: Introduction to Characterizing Binding via ITC

A Practical Approach to Microcalorimetry: Applications of Nano DSC

Instrument Information:

Waters offers microcalorimeters through its TA Instruments division:

Affinity ITC - Designed for the most challenging life science laboratory environments that require high sensitivity, high productivity and the most advanced ITC technologies. Nano DSC - Offering unmatched flexibility for characterizing molecular structure and stability.