Bujalowski Wlodzimierz, Jezewska Maria J, Bujalowski Paul J
Department of Biochemistry and Molecular Biology, The Sealy Center for Structural Biology, Sealy Center for Cancer Cell Biology, The University of Texas Medical Branch at Galveston, 301 University Boulevard, Galveston, TX 77555-1053, USA; Department of Obstetrics and Gynecology, The Sealy Center for Structural Biology, Sealy Center for Cancer Cell Biology, The University of Texas Medical Branch at Galveston, 301 University Boulevard, Galveston, TX 77555-1053, USA.
Department of Biochemistry and Molecular Biology, The Sealy Center for Structural Biology, Sealy Center for Cancer Cell Biology, The University of Texas Medical Branch at Galveston, 301 University Boulevard, Galveston, TX 77555-1053, USA.
Biophys Chem. 2017 Mar;222:25-40. doi: 10.1016/j.bpc.2016.12.005. Epub 2016 Dec 30.
Physico-chemical titration techniques are the most commonly used methods in characterizing molecular interactions. These methods are mainly based on spectroscopic, calorimetric, hydrodynamic, etc., measurements. However, truly quantitative physico-chemical methods are absolutely based on the determination of the relationship between the measured signal and the total average degree of binding in order to obtain meaningful interaction parameters. The relationship between the observed physico-chemical signal of whatever nature and the degree of binding must be determined and not assumed, based on some ad hoc intuitive relationship/model, leading to determination of the true binding isotherm. The quantitative methods reviewed and discussed here allow an experimenter to rigorously determine the degree of binding and the free ligand concentration, i.e., they lead to the construction of the thermodynamic binding isotherm in a model-independent fashion from physico-chemical titration curves.
物理化学滴定技术是表征分子相互作用时最常用的方法。这些方法主要基于光谱、量热、流体动力学等测量。然而，真正的定量物理化学方法绝对是基于确定测量信号与总平均结合度之间的关系，以便获得有意义的相互作用参数。必须确定而非基于某些临时的直观关系/模型假设任何性质的观测物理化学信号与结合度之间的关系，从而确定真实的结合等温线。这里回顾和讨论的定量方法使实验者能够严格确定结合度和游离配体浓度，即它们能够以与模型无关的方式从物理化学滴定曲线构建热力学结合等温线。
