Atomic absorption spectrometric detection of biotin-cymantrene as a metallo-tracer for the avidin-biotin system.
Biotin-cyclopentadienylmanganese tricarbonyl (biotin-cymantrene; biotin-Cy) is proposed as a universal metallo-tracer for immunoassays. Associated with the possibility of detecting this type of label (organometallic moieties) along with different analytical procedures, this reagent appears to be a universal system with the advantages of being inexpensive, stable and directly detected (no substrate is required). The optimum conditions for detecting the organometallic label biotin-Cy with a Zeeman atomic absorption spectrometer and the affinity of biotin-Cy for the streptavidin ligand immobilized on the walls of microtitre plates, wells or tubes are described. The results obtained demonstrate the possibility of assaying a wide range of clinical parameters because the detection limit of biotin-Cy is about 1 pg per assay. Based on the competition between immobilized bovine serum albumin adsorbed on the wall and free bovine serum albumin for specific polyclonal antibodies, the streptavidin-metallo-biotin system was tested on a competitive immunoassay model and a satisfactory calibration graph was obtained.
Rémy
I
I
Unité d'Immunoanalyse, Faculté de Pharmacie, Dijon, France.
Brossier
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P
D000818:Animals; D001360:Avidin; D001710:Biotin / Q000031:analogs & derivatives* / Q000032:analysis; D002417:Cattle; D007118:Immunoassay / Q000379:methods; D007700:Kinetics; D009942:Organometallic Compounds / Q000032:analysis*; D012709:Serum Albumin / Q000032:analysis*; D013054:Spectrophotometry, Atomic / Q000379:methods
D009942:Organometallic Compounds; D012709:Serum Albumin; D001360:Avidin; C083373:biotinylcyclopentadienylmanganese tricarbonyl; D001710:Biotin