The antigen-binding domains of camelid heavy-chain antibodies, also called nanobodies, gained strong attention because of their unique functional and biophysical properties. They gave rise to an entire spectrum of applications in biotechnology, research and medicine. Despite several reports about reversibly refolding nanobodies, protein aggregation plays a major role in nanobody thermoresistance, asking for strategies to engineer their refolding behavior. Here, we use measurements of nanobody aggregation kinetics to validate structural features in the nanobody fold that are suppressing heat-induced nanobody aggregation. Furthermore, the kinetic measurements yielded a detailed insight into the concept of the ΔTm shift, a metric for protein aggregation propensities obtained from differential scanning fluorimetry measurements. By relating the equilibrium measurements of the ΔTm shift to the kinetic measurements of heat-induced nanobody aggregation, a distinct relationship could be identified that allows a prediction of nanobody aggregation rates from a simple equilibrium measurement of ΔTm.

中文翻译：

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利用ΔTm位移进行纳米体稳定性工程；与热诱导聚集的表观速率常数的比较。

骆驼科重链抗体的抗原结合域（也称为纳米抗体）因其独特的功能和生物物理特性而受到了广泛的关注。他们带来了在生物技术，研究和医学中的整个应用领域。尽管有几篇有关可逆性折叠纳米抗体的报道，但蛋白质聚集在纳米抗体的热阻中起着重要作用，要求采取策略来设计其折叠行为。在这里，我们使用纳米抗体聚集动力学的测量结果来验证纳米抗体折叠中抑制热诱导纳米抗体聚集的结构特征。此外，动力学测量对ΔTm位移的概念有了更深入的了解，ΔTm位移是一种通过差示扫描荧光法测量获得的蛋白质聚集倾向的度量。