Development of binding assays in microfabricated picoliter vials: An assay for biotin

Anne L. Grosvenor; Agatha Feltus; Richard C. Conover; Sylvia Daunert; Kimberly W. Anderson

doi:10.1021/ac991289+

Development of binding assays in microfabricated picoliter vials: An assay for biotin

Anne L. Grosvenor, Agatha Feltus, Richard C. Conover, Sylvia Daunert, Kimberly W. Anderson

Research output: Contribution to journal › Article › peer-review

27 Scopus citations

Abstract

A homogeneous binding assay for the detection of biotin in picoliter vials was developed using the photoprotein aequorin as the label. The binding assay was based on the competition of free biotin with biotinylated aequorin (AEQ-biotin) for avidin. A sequential protocol was used, and modification of the assay to reduce the number of steps was examined. Results showed that detection limits on the order of 10^-14 mol of biotin were possible. Reducing the number of steps provided similar detection limits but only if the amount of avidin used was decreased. These binding assays based on picoliter volumes have potential applications in a variety of fields, including microanalysis and single-cell analysis, where the amount of sample is limited. In addition, these assays are suitable for the high-throughput screening of biopharmaceuticals.

Original language	English
Pages (from-to)	2590-2594
Number of pages	5
Journal	Analytical Chemistry
Volume	72
Issue number	11
DOIs	https://doi.org/10.1021/ac991289+
State	Published - Jun 1 2000

Funding

Funders	Funder number
National Institute of General Medical Sciences	R01GM047915

ASJC Scopus subject areas

Analytical Chemistry

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abstract = "A homogeneous binding assay for the detection of biotin in picoliter vials was developed using the photoprotein aequorin as the label. The binding assay was based on the competition of free biotin with biotinylated aequorin (AEQ-biotin) for avidin. A sequential protocol was used, and modification of the assay to reduce the number of steps was examined. Results showed that detection limits on the order of 10-14 mol of biotin were possible. Reducing the number of steps provided similar detection limits but only if the amount of avidin used was decreased. These binding assays based on picoliter volumes have potential applications in a variety of fields, including microanalysis and single-cell analysis, where the amount of sample is limited. In addition, these assays are suitable for the high-throughput screening of biopharmaceuticals.",

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T2 - An assay for biotin

AU - Grosvenor, Anne L.

AU - Feltus, Agatha

AU - Conover, Richard C.

AU - Daunert, Sylvia

AU - Anderson, Kimberly W.

PY - 2000/6/1

Y1 - 2000/6/1

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AB - A homogeneous binding assay for the detection of biotin in picoliter vials was developed using the photoprotein aequorin as the label. The binding assay was based on the competition of free biotin with biotinylated aequorin (AEQ-biotin) for avidin. A sequential protocol was used, and modification of the assay to reduce the number of steps was examined. Results showed that detection limits on the order of 10-14 mol of biotin were possible. Reducing the number of steps provided similar detection limits but only if the amount of avidin used was decreased. These binding assays based on picoliter volumes have potential applications in a variety of fields, including microanalysis and single-cell analysis, where the amount of sample is limited. In addition, these assays are suitable for the high-throughput screening of biopharmaceuticals.

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Development of binding assays in microfabricated picoliter vials: An assay for biotin

Abstract

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