Bioluminescence Binding Assay for Biotin with Attomole Detection Based on Recombinant Aequorin

Allan Witkowski, Sridhar Ramanathan, Sylvia Daunert

Research output: Contribution to journalArticle

36 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)1837-1840
Number of pages4
JournalAnalytical Chemistry
Volume66
Issue number11
DOIs
StatePublished - Jun 1 1994
Externally publishedYes

ASJC Scopus subject areas

  • Analytical Chemistry

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