Dual analyte detection using tandem flash luminescence.
A heterogeneous, dual analyte-binding assay which makes use of the flash luminescence from both aequorin and an acridinium-9-carboxamide label is presented. The signal generating species were triggered both differentially and sequentially using Ca(2+) followed by basic peroxide. Both signals were resolved readily using a single photomultiplier tube without the need for multiwavelength detection. To demonstrate the tandem luminescence concept in a model assay system, dose-response curves for two analytes, biotinylated BSA and myoglobin, were generated using a competitive binding format. Because of the relatively short assay time and the well-resolved signals, this format will be useful in the development of dual analyte high-throughput assays.
Adamczyk
Maciej
M
Department of Chemistry (9NM), Abbott Diagnostics Division, Abbott Laboratories, 60064-6016, Abbott Park, IL 60064-6016, USA. maciej.adamczyk@abbott.com
Moore
Jeffrey A
JA
Shreder
Kevin
K
D000166:Acridines / Q000737:chemistry*; D000331:Aequorin / Q000737:chemistry*; D000818:Animals; D001360:Avidin / Q000737:chemistry; D001667:Binding, Competitive; D002118:Calcium / Q000737:chemistry; D002417:Cattle; D004305:Dose-Response Relationship, Drug; D006801:Humans; D007202:Indicators and Reagents; D007700:Kinetics; D049449:Luminescence; D008954:Models, Biological; D009211:Myoglobin / Q000032:analysis; D011994:Recombinant Proteins / Q000032:analysis; D012710:Serum Albumin, Bovine / Q000032:analysis
D000166:Acridines; D007202:Indicators and Reagents; D009211:Myoglobin; D011994:Recombinant Proteins; C402305:acridinium-9-carboxamide; D001360:Avidin; D012710:Serum Albumin, Bovine; D000331:Aequorin; D002118:Calcium