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We compare aequorin, alkaline phosphatase and horseradish peroxidase as reporters for luminescent immunoassays using α-fetoprotein (AFP) as a model analyte. Biotinylated aequorin was prepared from mutated aequorin containing a reactive cysteine residue by chemical conjugation. The measurable range of AFP using this biotinylated aequorin was 0.02–200 ng/ml, with a lower background level than the other biotinylated enzymes tested.
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