Application of a stepwise frontal analysis method in cell membrane chromatography

J Chromatogr B Analyt Technol Biomed Life Sci. 2020 Dec 15:1161:122436. doi: 10.1016/j.jchromb.2020.122436. Epub 2020 Nov 18.

Abstract

Bio-affinity chromatography is used in the study of drug-receptor interactions. A stepwise frontal analysis (SFA) method was developed based on frontal analysis (FA). A high expression alpha 1A adrenergic receptor (α1A AR) cell membrane chromatography (CMC) method was then developed and combined with SFA to investigate the affinity of three model α1A AR-binding drugs towards α1A AR. Equilibrium dissociation constant (Kd) values for drug-receptor interactions were determined by FA and SFA; results showed that these methods were highly consistent. The results demonstrate that the CMC/SFA method is a time-saving and less wasteful method than traditional method for the evaluation of drug-receptor binding characteristics, and could be used to study the interactions between drugs and membrane receptors.

Keywords: Cell membrane chromatography; Equilibrium dissociation constant (K(d)); Stepwise frontal analysis.

MeSH terms

  • Cell Membrane / metabolism*
  • Chromatography, Affinity / methods*
  • Drug Discovery
  • HEK293 Cells
  • Humans
  • Pharmaceutical Preparations / analysis
  • Pharmaceutical Preparations / chemistry
  • Pharmaceutical Preparations / metabolism
  • Protein Binding
  • Receptors, Adrenergic, alpha-1 / metabolism

Substances

  • Pharmaceutical Preparations
  • Receptors, Adrenergic, alpha-1