Durowoju Ibrahim B, Bhandal Kamaljit S, Hu Jian, Carpick Bruce, Kirkitadze Marina
Analytical Research & Development, Sanofi Pasteur Limited.
Analytical Research & Development, Sanofi Pasteur Limited;
J Vis Exp. 2017 Mar 4(121):55262. doi: 10.3791/55262.
Differential scanning calorimetry (DSC) is an analytical technique that measures the molar heat capacity of samples as a function of temperature. In the case of protein samples, DSC profiles provide information about thermal stability, and to some extent serves as a structural "fingerprint" that can be used to assess structural conformation. It is performed using a differential scanning calorimeter that measures the thermal transition temperature (melting temperature; Tm) and the energy required to disrupt the interactions stabilizing the tertiary structure (enthalpy; ∆H) of proteins. Comparisons are made between formulations as well as production lots, and differences in derived values indicate differences in thermal stability and structural conformation. Data illustrating the use of DSC in an industrial setting for stability studies as well as monitoring key manufacturing steps are provided as proof of the effectiveness of this protocol. In comparison to other methods for assessing the thermal stability of protein conformations, DSC is cost-effective, requires few sample preparation steps, and also provides a complete thermodynamic profile of the protein unfolding process.
差示扫描量热法（DSC）是一种分析技术，用于测量样品的摩尔热容随温度的变化。对于蛋白质样品，DSC曲线可提供有关热稳定性的信息，并在一定程度上作为一种结构“指纹”，可用于评估结构构象。该技术使用差示扫描量热仪进行，可测量蛋白质的热转变温度（熔化温度；Tm）以及破坏稳定三级结构的相互作用所需的能量（焓；∆H）。可对不同配方以及不同生产批次进行比较，所得值的差异表明热稳定性和结构构象存在差异。文中提供了说明DSC在工业环境中用于稳定性研究以及监测关键生产步骤的数据，以证明该方案的有效性。与其他评估蛋白质构象热稳定性的方法相比，DSC具有成本效益，所需的样品制备步骤较少，还能提供蛋白质展开过程的完整热力学曲线。
