The Power of Heat: Digging Deeper with Differential Scanning Calorimetry to Study Key Protein Characteristics

Differential Scanning Calorimetry (DSC) has a long history of playing a vital role in biopharmaceutical development. From candidate selection to formulation development, stability monitoring and bio-similarity assessment, DSC remains one of the top assays utilized in the pharmaceutical industry during the development and approval of protein-based therapeutics.

There is familiarity in the biopharmaceutical industry with the basic concept of DSC as it relates to protein stability. As a protein is heated in a DSC experiment, the midpoint of thermal denaturation transition(s) (termed Tm) is/are measured, and ranking of stability based on higher Tm values is performed (whether different constructs or different formulations of a protein, etc.). Although extremely useful for protein stability assessment, Tm values alone may not always reveal some characteristics of a protein molecule that can be highly dependent on the solution environment.

DSC analyses can be used to probe even deeper when other parameters obtained from the assay are also used (onset temperature of unfolding, peak width and peak area), even when it appears that a Tm does not shift. In this presentation, real-life examples of taking DSC analysis “deeper” will be presented. These examples will include monitoring the binding of a metal ion to an enzyme, high(er) throughput formulation development using a DOE approach for a monoclonal antibody, and using DSC to assist in the development of a complex purification process for a fusion protein.