title: Development and characterization of immobilized cannabinoid receptor (CB1/CB2) open tubular column for on-line screening.
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.
authors.affiliation: Gerontology Research Center, National Institute on Aging, National Institutes of Health, Baltimore, MD 21224, USA. moaddelru@grc.nia.nih.gov
authors.forename: R
authors.initials: R
authors.lastname: Moaddel
authors.forename: A
authors.initials: A
authors.lastname: Rosenberg
authors.forename: K
authors.initials: K
authors.lastname: Spelman
authors.forename: J
authors.initials: J
authors.lastname: Frazier
authors.forename: C
authors.initials: C
authors.lastname: Frazier
authors.forename: S
authors.initials: S
authors.lastname: Nocerino
authors.forename: A
authors.initials: A
authors.lastname: Brizzi
authors.forename: C
authors.initials: C
authors.lastname: Mugnaini
authors.forename: I W
authors.initials: IW
authors.lastname: Wainer
mesh_terms: D002186:Cannabinoids / Q000737:chemistry; D045744:Cell Line, Tumor; D002846:Chromatography, Affinity / Q000379:methods*; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry; D018517:Plant Roots / Q000737:chemistry; D011485:Protein Binding; D043884:Receptor, Cannabinoid, CB1 / Q000819:agonists / Q000737:chemistry*; D043885:Receptor, Cannabinoid, CB2 / Q000819:agonists / Q000737:chemistry*; D032106:Zanthoxylum / Q000737:chemistry
chemical_list: D002186:Cannabinoids; D055767:Immobilized Proteins; D043884:Receptor, Cannabinoid, CB1; D043885:Receptor, Cannabinoid, CB2