I would like to fit a curve of my steady-state anisotropy results in Origin, which revealed weak binding between two proteins. I tried quadratic equation but it turned out that it is used for strong interction, so I would like to reevaluate my results. I tried to find information about weak binding fits, but could not find detailed infos.
Thank you in advance!
I suggest you follow https://shoichetlab.utoronto.ca/wp-content/uploads/2018/07/mathematical-model...affinity-based-delivery-system.pdf
Article Mathematical Model of Dynamic Protein Interactions Regulatin...
See some hints/tips on these posts:
https://www.researchgate.net/post/Calculating_kd_dissociation_constant_using_Prism
https://www.researchgate.net/post/How_can_I_estimate_Kd_value_of_protein-protein_interactions_If_I_have_measurements_only_in_linear_range
The general quadratic binding equation (relating total protein concentrations, dissociation constant, and saturation fraction) is valid for any binding affinity (low, moderate, and high).
However, if affinity is low and, importantly, the concentration of the titrating protein in all experimental points of the binding isotherm is much higher than that of the titrated protein, you may use a binding model based on a hyperbolic function (equivalent to Langmuir model), which is easier to handle, but it involves an approximation (no depletion of titrating protein concentration because of binding and equivalence of total concentration and free concentration)
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