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 journalArticle

25 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 languageEnglish (US)
Pages (from-to)2590-2594
Number of pages5
JournalAnalytical Chemistry
Volume72
Issue number11
DOIs
StatePublished - Jun 1 2000
Externally publishedYes

ASJC Scopus subject areas

  • Analytical Chemistry

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