Abstract

Formulation conditions have a significant influence on the degree of freeze/thaw (FT) stress-induced protein instabilities. Adding cryoprotectants might stabilize the induced FT stress instabilities. However, a simple preservation of protein stability might be insufficient and further methods are necessary. This study aims to evaluate the addition of a heat cycle following FT application as a function of different cryoprotectants with lysozyme as exemplary protein. Sucrose and glycerol were shown to be the most effective cryoprotectants when compared to PEG200 and Tween20. In terms of heat-induced reversibility of aggregates, glycerol showed the best performance followed by sucrose, NaCl and Tween20 systems. The analysis was performed using a novel approach to visualize complex interplays by a clustering and data reduction scheme. In addition, solubility and structural integrity were measured and confirmed the obtained results.

Graphic abstract

中文翻译：

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使用热循环作为不同防冻剂的函数研究冷冻/解冻应力诱导的蛋白质不稳定性的可逆性。

摘要

配制条件对冷冻/解冻（FT）应激诱导的蛋白质不稳定性程度具有重大影响。添加防冻剂可能会稳定所引起的FT应力不稳定性。然而，简单地保持蛋白质稳定性可能是不够的，并且需要进一步的方法。这项研究的目的是根据溶菌酶作为示例性蛋白质，评估不同的冷冻保护剂对FT应用后热循环的增加。与PEG200和Tween20相比，蔗糖和甘油被证明是最有效的防冻剂。就聚集体的热诱导可逆性而言，甘油表现出最佳性能，其次是蔗糖，NaCl和Tween20系统。使用新颖的方法进行分析，以通过聚类和数据缩减方案可视化复杂的相互作用。此外，

图形摘要