Development of a radioimmunoassay for idazoxan hydrochloride.
To facilitate the measurement of idazoxan in clinical trials an immunoassay capable of detecting low ng ml-1 concentrations was required. A stable derivative was prepared which, after linking to keyhole limpet hemocyanin (KLH), was subsequently immunized into sheep. Early assay development was carried out with both fluorescent (1-FITC) and iodinated (I-125) labels. The assay methodology was the same in both cases, using magnetizable solid-phase particles to which the antibody was linked. The sensitivity of the assay was such that a large sample volume was required, which in turn, led to problems of increased protein interference. The use of pepsin to digest the protein was used effectively after several blocking agents were unsuccessfully used. The limit of detection was in the region of 3 ng ml-1. Cross-reactivity studies showed that the antibody was specific for idazoxan. Intra- and inter-assay precision was 7 and 12%, respectively. Correlation with the analytical GC-MSD method was in the order of 0.90.
Lynn
A G
AG
Analytical Services Department, Reckitt and Colman Pharmaceuticals, Hull, North Humberside, UK.
Coxon
R E
RE
Gallacher
G
G
Landon
J
J
D000918:Antibody Specificity; D003429:Cross Reactions; D004146:Dioxanes / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D016231:Fluorescence Polarization Immunoassay; D005455:Fluorescent Antibody Technique; D006433:Hemocyanins; D006801:Humans; D019329:Idazoxan; D011863:Radioimmunoassay
D004146:Dioxanes; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin; D019329:Idazoxan