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

Sylvia Daunert; Leonidas G. Bachas; Mark E. Meyerhoff

doi:10.1016/S0003-2670(00)80734-5

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 journal › Article › peer-review

29 Scopus citations

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 language	English (US)
Pages (from-to)	43-52
Number of pages	10
Journal	Analytica Chimica Acta
Volume	208
Issue number	C
DOIs	https://doi.org/10.1016/S0003-2670(00)80734-5
State	Published - 1988
Externally published	Yes

ASJC Scopus subject areas

Analytical Chemistry
Biochemistry
Environmental Chemistry
Spectroscopy

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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.",

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AU - Daunert, Sylvia

AU - Bachas, Leonidas G.

AU - Meyerhoff, Mark E.

PY - 1988

Y1 - 1988

N2 - 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.

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