Unlock Exclusive Discounts & Flash Sales! Click Here to Join the Deals on Every Wednesday!

Analysis of Conformational Stability of Proteins by Differential Scanning Calorimetry (µDSC) (CAT#: STEM-B-0372-CJ)

Introduction

Structure and conformation of a biological molecule is key for its function. The higher order structure of a biopharmaceutical molecule is, thereby, often directly connected to the quality, stability, safety, and efficacy of a therapy. The higher order structure is considered a critical quality attribute and, thus, a detailed understanding of the higher order structure of a biopharmaceutical compound is critical in every research and development phase. Characterizing the secondary, tertiary and, if present, quaternary structure of a biopharmaceutical compound requires multiple analytical techniques.




Principle

During µDSC analysis, the protein of interest is exposed to a temperature ramp, where the required thermal energy for each temperature increase is recorded in correlation to a reference sample (e.g., formulation buffer). As a result, protein unfolding events can be observed and the melting temperature (Tm) of each event can be determined. Also, the unfolding enthalpy (ΔH) of the protein or individual protein domains can be calculated from the area under the melting curve. µDSC is also employed to study the reversibility of unfolding events and can give insight into higher-order structures. µDSC is considered an “absolute technique”, as it does not rely on any reference standards to obtain quantitative results. However, µDSC requires much more time to analyze one sample than the orthogonal technique nanoDSF.

Applications

Biopharmaceutica

Procedure

1. Preliminaries.
2. Turn on the Cooling Unit.
3. Weigh Out the Sample.
4. Crimp the Sample.
5. Load the Sample Into the DSC Machine.
6. Prepare the Program.
7. Set Up DSC Machine.
8. Start the Analysis.

Materials

• Sample: Proteins
• Equipment: Differential scanning calorimetry (µDSC)

Notes

µDSC is an important and frequently used technique for the determination of the thermal stability of proteins.
Advertisement