Accurate titration of avidin and streptavidin with biotin–fluorophore conjugates in complex, colored biofluids

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• 作者：Gruber ; Hermann J. ; Kada ; Gerald ; Marek ; Markus ; Kaiser ; Karl
• 关键词：Avidin ; Biotin ; Fluorescein ; Fluorescence ; Streptavidin ; ANS ; 2-anilinonaphthalene-6-sulfonic acid ; Biotin&ndash ; PEG₈₀₀&ndash ; pyrene ; N-biotinoyl-N&prime ; -(1-pyrene)butanoyl-O ; O&prime ; -bis(2-aminopropyl)-poly(ethylene glycol)₈₀₀ ; EDT
• 刊名：Biochimica et Biophysica Acta (BBA)/General Subjects
• 出版年：1998
• 出版时间：July 23, 1998
• 年：1998
• 卷：1381
• 期：2
• 页码：203-212
• 全文大小：275 K

文摘

A new fluorimetric assay is presented for the specific and reliable quantitation of Ȧ5;2 nM avidin and streptavidin. The assay is based on pronounced changes in the fluorescence properties of commercial fluorescein–biotin, or of a newly synthesized biotin–poly(ethylene glycol)–pyrene conjugate, which occur upon binding to avidin and streptavidin. Accurate measurement of (strept)avidin in complex, colored biofluids, such as crude egg white or serum relies on a simple titration protocol. Only occasional recalibration of the reagent solution is required. Due to these merits the proposed assay is particularly suited for rapid measurement of few samples on short notice, for functional control of (strept)avidin-containing reagents after storage, and for the monitoring of (strept)avidin concentrations in large scale processes.