Formulation and Analytical Resources, Amgen Inc., Seattle, Washington, USA.
J Pharm Sci. 2010 Apr;99(4):1707-20. doi: 10.1002/jps.21955.
Differential scanning fluorimetry (DSF) was employed to increase the throughput of the thermostability screening of monoclonal antibody (mAb) formulations. The method consists of measuring the fluorescence intensity of a polarity sensitive probe at gradually increasing temperatures, and obtaining the transition temperature of exposure of the hydrophobic regions of proteins (T(h)). The change in fluorescence intensity was directly related to protein unfolding levels and temperatures. The results from thermostability measurements were compared with the data acquired using differential scanning calorimetry (DSC), and a good correlation between T(h) and the temperature of protein unfolding or melting (T(m)) was observed. The method was applied to screen four mAb molecules in 84 different buffers. The studies revealed a good correlation of T(h) values with the known effects of pH and excipients on protein stability in solution. Specifically, the elevated aggregation levels induced by salt, low pH, and high protein concentrations could be successfully predicted by this thermal stability screening. This method is efficient, with high throughput capability, and could be widely applied in the biopharmaceutical industry for formulation and process development, and characterization.
差示扫描荧光法（DSF）被用于提高单克隆抗体（mAb）制剂热稳定性筛选的通量。该方法包括测量在逐渐升高的温度下对极性敏感探针的荧光强度，并获得蛋白质疏水区暴露的转变温度（T(h)）。荧光强度的变化与蛋白质的展开水平和温度直接相关。热稳定性测量的结果与使用差示扫描量热法（DSC）获得的数据进行了比较，并观察到 T(h)与蛋白质展开或熔化温度（T(m)）之间存在良好的相关性。该方法应用于在 84 种不同缓冲液中筛选四种 mAb 分子。研究表明，T(h)值与 pH 值和赋形剂对溶液中蛋白质稳定性的已知影响之间存在良好的相关性。具体来说，通过这种热稳定性筛选可以成功预测盐、低 pH 值和高蛋白质浓度引起的聚集水平升高。该方法高效，具有高通量能力，可广泛应用于生物制药行业的制剂和工艺开发以及特性研究。
