Overview of the detection methods for equilibrium dissociation constant KD of drug-receptor interaction

Abstract

Drug-receptor interaction plays an important role in a series of biological effects, such as cell proliferation, immune response, tumor metastasis, and drug delivery. Therefore, the research on drug-receptor interaction is growing rapidly. The equilibrium dissociation constant (KD) is the basic parameter to evaluate the binding property of the drug-receptor. Thus, a variety of analytical methods have been established to determine the KD values, including radioligand binding assay, surface plasmon resonance method, fluorescence energy resonance transfer method, affinity chromatography, and isothermal titration calorimetry. With the invention and innovation of new technology and analysis method, there is a deep exploration and comprehension about drug-receptor interaction. This review discusses the different methods of determining the KD values, and analyzes the applicability and the characteristic of each analytical method. Conclusively, the aim is to provide the guidance for researchers to utilize the most appropriate analytical tool to determine the KD values.

The anti‐anaphylactoid effects of Piperine through regulating MAS‐related G protein‐coupled receptor X2 activation

“…CMC method is an effective technique to study the characteristics of drug‐membrane receptor affinity, which can be used to determine the K D value of drug‐membrane receptor (Ma et al, ; Ma, Yang, & He, ). For this experiment, CMC analysis was performed on a Shimadzu LC‐20A apparatus that consisted of two LC‐20AD pumps, a DGU‐20A3 degasser, a SIL‐20A auto sampler, a CTO‐20A column oven, and a SPD‐M20A diode array detector (Shimadzu, Kyoto, Japan).…”

“…In the CMC model, the K D value is investigated by using frontal analysis and the relative K D value as the following equation (Ma et al, ; Ma et al, ): where k is the capacity factor of the applied analyte, K DA is the equilibrium dissociation constant of the known analyte, K DI is the dissociation constant of the applied analyte, [ A ] is the molar concentration of applied analyte, m L is the total moles of binding sites in the column, V M is the dead volume of the CMC. In the CMC results, the k value could be calculated by the equation, k = ( t R − t 0 )/ t 0 , where t 0 is the retention time of solvent, and t R is the retention time of the known analyte.…”

Metal Chelation Therapy and Parkinson’s Disease: A Critical Review on the Thermodynamics of Complex Formation between Relevant Metal Ions and Promising or Established Drugs

“…However, as the proton content is experimentally not controlled when K d values are measured, not only for the ligand but also for the metal ion, we think that Equation 7allows a more rigorous calculation of K d values. Literature appears to have preference for measuring and reporting K d more than K a [313,315]. This is probably due to the chemical usefulness and significance of K d , as it represents the concentration of free metal ion at which the concentrations of free ligand and of the complexes are equivalent [313].…”

“…Values of K d can be determined with a much simpler experimental design than that used to obtain logβ, logK and pM [313,315]. This is a crucial advantage when complicated ligands are studied, such as proteins, for which the determination of logβ or logK is practically impossible.…”

Recent progress in the molecular imaging of therapeutic monoclonal antibodies

“…Assessment of antibody-antigen interaction is therefore important for understanding the efficacy and toxicity of mAb or immunoconjugate treatment [ 83 ]. In vitro binding affinity of mAbs can be detected by methods as described for serum mAb concentrations including ELISA [ 84 ] and SPR assay [ 85 , 86 ], so they will not be specified here. Very recently, Yao’s group [ 87 ] reported a sensor based on photonic crystal hydrogels (PCHs), which is capable of naked-eye detection of antibody-antigen interaction with high sensitivity, providing inspiration for detecting the interaction with advanced materials.…”