Departamento de Física de Aplicada, Facultade de Física, Universidade de Santiago de Compostela, E-15782, Santiago de Compostela, Spain.
AFFINImeter Scientific & Development team, Software 4 Science Developments, S. L. Ed. Emprendia, Santiago de Compostela, A Coruña, 15782, Spain.
Anal Biochem. 2019 Jul 15;577:117-134. doi: 10.1016/j.ab.2019.02.031. Epub 2019 Mar 5.
The comprehension of molecular recognition phenomena demands the understanding of the energetic and kinetic processes involved. General equations valid for the thermodynamic analysis of any observable that is assessed as a function of the concentration of the involved compounds are described, together with their implementation in the AFFINImeter software. Here, a maximum of three different molecular species that can interact with each other to form an enormous variety of supramolecular complexes are considered. The corrections currently employed to take into account the effects of dilution, volume displacement, concentration errors and those due to external factors, especially in the case of ITC measurements, are included. The methods used to fit the model parameters to the experimental data, and to generate the uncertainties are described in detail. A simulation tool and the so called kinITC analysis to get kinetic information from calorimetric experiments are also presented. An example of how to take advantage of the AFFINImeter software for the global multi-temperature analysis of a system exhibiting cooperative 1:2 interactions is presented and the results are compared with data previously published. Some useful recommendations for the analysis of experiments aimed at studying molecular interactions are provided.
对分子识别现象的理解需要了解所涉及的能量和动力学过程。本文描述了适用于作为所涉及化合物浓度函数评估的任何可观察量的热力学分析的通用方程，并介绍了它们在 AFFINImeter 软件中的实现。在这里，最多考虑三种不同的分子物种，它们可以相互作用形成种类繁多的超分子配合物。目前采用的修正方法包括考虑稀释、体积位移、浓度误差以及外部因素的影响，特别是在 ITC 测量的情况下。详细描述了用于将模型参数拟合到实验数据并生成不确定度的方法。还介绍了一种模拟工具和所谓的 kinITC 分析，以从量热实验中获取动力学信息。本文还展示了如何利用 AFFINImeter 软件对表现出协同 1:2 相互作用的系统进行全局多温度分析的示例，并将结果与先前发表的数据进行比较。提供了一些有用的建议，用于分析旨在研究分子相互作用的实验。
