Differential Scanning Calorimetry to Quantify Heat-Induced Aggregation in Concentrated Protein Solutions

  • Matthew R. Jacobs
  • Mark Grace
  • Alice Blumlein
  • Jennifer J. McManusEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2039)


Differential scanning calorimetry (DSC) is an important technique to measure the thermodynamics of protein unfolding (or folding). Information including the temperature for the onset of unfolding, the melt transition temperature (Tm), enthalpy of unfolding (ΔH), and refolding index (RI) are useful for evaluating the heat stability of proteins for a range of biochemical, structural biology, industrial, and pharmaceutical applications. We describe a procedure for careful sample preparation of proteins for DSC measurements and data analysis to determine a range of thermodynamic parameters. In particular, we highlight a measure of protein refolding following complete thermal denaturation (RI), which quantifies the proportion of protein lost to irreversible aggregation after thermal denaturation.

Key words

Differential scanning calorimetry Melt transition temperature Enthalpy Refolding index Protein aggregation Thermal denaturation 



The research work presented in this chapter was made possible through the financial support of Enterprise Ireland under grant number [IP/2015 0358] and the Synthesis and Solid State Pharmaceutical Centre, cofunded under the European Regional Development Fund, and the Department of Agriculture Food and Marine, FIRM grant number [11/F/037], SFI Stokes Lectureship to J.J.M., and Maynooth University Teaching Fellowship to A.B.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Matthew R. Jacobs
    • 1
  • Mark Grace
    • 1
  • Alice Blumlein
    • 1
  • Jennifer J. McManus
    • 2
    Email author
  1. 1.Department of ChemistryMaynooth UniversityMaynoothIreland
  2. 2.Department of ChemistryMaynooth UniversityMaynoothIreland

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