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

Matthew R Jacobs; Mark Grace; Alice Blumlein; Jennifer J McManus

doi:10.1007/978-1-4939-9678-0_9

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

Methods Mol Biol. 2019:2039:117-129. doi: 10.1007/978-1-4939-9678-0_9.

Authors

Matthew R Jacobs¹, Mark Grace¹, Alice Blumlein¹, Jennifer J McManus²

Affiliations

¹ Department of Chemistry, Maynooth University, Maynooth, Kildare, Ireland.
² Department of Chemistry, Maynooth University, Maynooth, Co. Kildare, Ireland. jennifer.mcmanus@mu.ie.

PMID: 31342423
DOI: 10.1007/978-1-4939-9678-0_9

Abstract

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 (T_m), 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.

Keywords: Differential scanning calorimetry; Enthalpy; Melt transition temperature; Protein aggregation; Refolding index; Thermal denaturation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Calorimetry, Differential Scanning / methods
Hot Temperature
Protein Aggregates / physiology
Protein Denaturation
Protein Folding
Proteins / chemistry*
Thermodynamics

Substances

Protein Aggregates
Proteins