Thermal stability is a critical quality attribute that is crucial to monitor during the development of therapeutic proteins to ensure their structural integrity and efficacy during manufacturing, storage and administration processes. Maintaining stability safeguards against denaturation or degradation, preserving the protein's therapeutic activity and enhancing patient safety and treatment outcomes.
This infographic compares four different methods to assess thermal stability and the unique value each provides.
Download this infographic to discover a method that:
- Is highly precise
- Requires low sample volume and is label-free
- Provides insights into melting temperature, onset of melting and energy of activation of unfolding
Four methods to assess the thermal stability of your therapeutic proteins How it’s done What it tells you Benefits Limitations Method nanoDSF Intrinsic Differential Scanning Fluorimetry CD Circular Dichroism Extrinsic DSF Thermofluor Differential Scanning Fluorimetry Follows shift in emission maximum from a protein’s intrinsic fluorescence to determine unfolding Melting temperature (Tm ) Onset of melting (Ton ) Energy of activation of unfolding (Ea ) Highly precise Tm measurement Low sample volume High throughput Label-free No secondary structure info Compare changes in polarized light output from sample in response to temperature gradient Melting temperature (Tm ) Enthalpy of unfolding (ΔH) Which secondary structure elements your protein contains Provides insight into secondary structure Low sample volume requirements Medium throughput Low buffer flexibility Measurements take some time to collect Fluorescent dye binds hydrophobic patches of protein as it unfolds in response to temperature denaturation Melting temperature (Tm ) Inexpensive, no special equipment Low sample volume requirements High throughput Difficult to discern multiple transitions Low buffer flexibility False stabilization from dye No secondary structure info DSC Differential Scanning Calorimetry Compares the difference in heat capacity between a sample and a reference Melting temperature (Tm ) Enthalpy of unfolding (ΔH) Highly precise Tm measurement Label-free Low throughput High sample consumption No secondary structure info In this table, learn about 4 different methods for thermal stability assessment of therapeutic proteins. While each provides unique value, the table will help you understand why nanoDSF is the go-to choice for determining the Tm s of your therapeuti