Streptavidin-Biotin Based Bioluminescent Enzyme Immunoassay Using Biotinylated Acetate Kinase and Recombinant Firefly Luciferase

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Abstract
Streptavidin-biotin system has been introduced to a bioluminescent enzyme immunoassay using biotinylated acetate kinase and recombinant firefly luciferase. The streptavidin-biotin based bioluminescent enzyme immunoassays (BLEIAs) for thyroid stimulating hormone (TSH) and human chorionic gonadotropin (hCG) were developed in which the complexes of biotinylated acetate kinase, streptavidin, and biotinylated antibody were used. In BLEIA for TSH, acetate kinase activity was determined by endpoint assay and for hCG the enzyme activity was measured by rate assay. The measurable ranges of TSH and hCG were 0.02 μIU/ml to 50 μIU/ml and 0.003 to 2 μIU/ml, respectively. The proposed streptavidin-biotinylated acetate kinase system was shown to be applicable to several sandwich type enzyme immunoassays employing biotinylated antibody with high sensitivity.