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 |
|---|---|
| Pages (from-to) | 43-52 |
| Number of pages | 10 |
| Journal | Analytica Chimica Acta |
| Volume | 208 |
| Issue number | C |
| DOIs | |
| State | Published - Jan 1 1988 |
| Externally published | Yes |
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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 journal › Article
}
TY - JOUR
T1 - Homogeneous enzyme-linked competitive binding assay for biotin based on the avidin-biotin interaction
AU - Daunert, Sylvia
AU - Bachas, Leonidas G
AU - Meyerhoff, Mark E.
PY - 1988/1/1
Y1 - 1988/1/1
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.
AB - 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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U2 - 10.1016/S0003-2670(00)80734-5
DO - 10.1016/S0003-2670(00)80734-5
M3 - Article
AN - SCOPUS:0023923788
VL - 208
SP - 43
EP - 52
JO - Analytica Chimica Acta
JF - Analytica Chimica Acta
SN - 0003-2670
IS - C
ER -