Processing of the spectrofluorimetric data using the graphical methods and the maximum likelihood approach

The present work describes the calculation of the binding constants from spectrofluorimetric data using simple graphical methods and specialized software implementing the maximum likelihood approach. The following popular cases are analyzed: 1) protein-small molecule; 2) protein-metal complex; 3) DNA-small molecule; 4) DNA-metal complex interactions. The inability of graphical plots to return the correct results except for the simplest situation (single reaction with a non-fluorescent product) is demonstrated. The possibility of determining the most probable stoichiometric model using the maximum likelihood estimation (LSQ as its special case) is discussed as well as the limitations. (C) 2020 Elsevier B.V. All rights reserved.

Automated summary

This study describes the calculation of binding constants from spectrofluorimetric data using simple graphical methods and specialized software implementing the maximum likelihood approach. The inability of graphical plots to return correct results except for the simplest situations is demonstrated. The possibility of determining the most probable stoichiometric model using maximum likelihood estimation is discussed, along with its limitations.