Biotinylated recombinant aequorin was used in the development of a homogeneous bioluminescence assay for biotin. It was demonstrated that the Ca2+-triggered luminescent reaction of biotinylated aequorin can be inhibited by the presence of avidin, and this inhibition in bioluminescence intensity can be used to determine the biotin concentration in a sample. The optimized assay thus developed had a detection limit for biotin of 4 amol (equivalent to 1 × 10-14 M in the assay system), which to our knowledge is the lowest detection limit reported to date for a homogeneous competitive binding assay.
ASJC Scopus subject areas
- Analytical Chemistry