Development and characterization of immobilized cannabinoid receptor (CB1/CB2) open tubular column for on-line screening

Anal Biochem. 2011 May 1;412(1):85-91. doi: 10.1016/j.ab.2010.12.034. Epub 2011 Jan 6.

Abstract

Cannabinoid receptors, CB1 and CB2, are therapeutic targets in the treatment of anxiety, obesity, movement disorders, glaucoma, and pain. We have developed an on-line screening method for CB1 and CB2 ligands, where cellular membrane fragments of a chronic myelogenous leukemia cell line, KU-812, were immobilized onto the surface of an open tubular (OT) capillary to create a CB1/CB2-OT column. The binding activities of the immobilized CB1/CB2 receptors were established using frontal affinity chromatographic techniques. This is the first report that confirms the presence of functional CB1 and CB2 receptors on KU-812 cells. The data from this study confirm that the CB1/CB2-OT column can be used to determine the binding affinities (K(i) values) for a single compound and to screen individual compounds or a mixture of multiple compounds. The CB1/CB2-OT column was also used to screen a botanical matrix, Zanthoxylum clava-herculis, where preliminary results suggest the presence of a high-affinity phytocannabinoid.

Publication types

  • Research Support, N.I.H., Intramural

MeSH terms

  • Cannabinoids / chemistry
  • Cell Line, Tumor
  • Chromatography, Affinity / methods*
  • Humans
  • Immobilized Proteins / chemistry
  • Plant Roots / chemistry
  • Protein Binding
  • Receptor, Cannabinoid, CB1 / agonists
  • Receptor, Cannabinoid, CB1 / chemistry*
  • Receptor, Cannabinoid, CB2 / agonists
  • Receptor, Cannabinoid, CB2 / chemistry*
  • Zanthoxylum / chemistry

Substances

  • Cannabinoids
  • Immobilized Proteins
  • Receptor, Cannabinoid, CB1
  • Receptor, Cannabinoid, CB2