Homogeneous enzyme-linked competitive binding assay for biotin based on the avidin-biotin interaction

Sylvia Daunert, Leonidas G Bachas, Mark E. Meyerhoff

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

A homogeneous enzyme-linked competitive-binding assay for biotin with glucose-6-phosphate dehydrogenase (G6PDH), is described. This assay is based on the interaction between a G6PDH/biotin conjugate with avidin, a natural binder for biotin. In the absence of biotin in the assay mixture, this interaction results in 100% inhibition of the enzyme conjugate. In the presence of biotin, the enzymatic activity of the conjugate is regained in an amount related to the concentration of the vitamin in the sample. Extremely steep, gate-like dose/response curves, attributable to the relative binding affinities of avidin for biotin and the conjugate, are observed. The detection limits of the system vary with the amounts of avidin and enzyme/biotin conjugate used. The method is rapid and sensitive and is evaluated for the direct determination of biotin in vitamin tablets.

Original languageEnglish
Pages (from-to)43-52
Number of pages10
JournalAnalytica Chimica Acta
Volume208
Issue numberC
DOIs
StatePublished - Jan 1 1988
Externally publishedYes

Fingerprint

Competitive Binding
Avidin
Biotin
Assays
assay
enzyme
vitamin
glucose
Enzymes
phosphate
Glucosephosphate Dehydrogenase
Vitamins
Tablets
Binders
Limit of Detection

ASJC Scopus subject areas

  • Biochemistry
  • Analytical Chemistry
  • Spectroscopy
  • Environmental Chemistry

Cite this

Homogeneous enzyme-linked competitive binding assay for biotin based on the avidin-biotin interaction. / Daunert, Sylvia; Bachas, Leonidas G; Meyerhoff, Mark E.

In: Analytica Chimica Acta, Vol. 208, No. C, 01.01.1988, p. 43-52.

Research output: Contribution to journalArticle

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