Sugar additives improve signal fidelity for implementing two-phase resorufin-based enzyme immunoassays

Enzymatic signal amplification based on fluorogenic substrates is commonly used for immunoassays; however, when transitioning these assays to a digital format in water-in-mineral oil emulsions, such amplification methods have been limited by the leakage of small reporting fluorescent probes. In the...

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Bibliographische Detailangaben
Veröffentlicht in:Langmuir : the ACS journal of surfaces and colloids. - 1992. - 30(2014), 23 vom: 17. Juni, Seite 6637-43
1. Verfasser: Sandoz, Patrick A (VerfasserIn)
Weitere Verfasser: Chung, Aram J, Weaver, Westbrook M, Di Carlo, Dino
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2014
Zugriff auf das übergeordnete Werk:Langmuir : the ACS journal of surfaces and colloids
Schlagworte:Journal Article Research Support, U.S. Gov't, Non-P.H.S. Micelles Oxazines Surface-Active Agents Water 059QF0KO0R resorufin 635-78-9
Beschreibung
Zusammenfassung:Enzymatic signal amplification based on fluorogenic substrates is commonly used for immunoassays; however, when transitioning these assays to a digital format in water-in-mineral oil emulsions, such amplification methods have been limited by the leakage of small reporting fluorescent probes. In the present study, we used a microfluidic system to study leakage from aqueous droplets in a controlled manner and confirmed that the leakage of fluorescent resorufin derivatives is mostly due to the presence of the lipophilic surfactant Span80, which is commonly used to preserve emulsion stability. This leakage can be overcome by the addition of specific sugars that most strongly interfered with the surfactants ability to form micelles in water. The application of the microfluidic system to the quantitative analysis of droplets and the implementation of the described sugar additives would allow for alternatives to fluorinated surfactant-based platforms and improve the signal fidelity in enzyme immunoassays implemented through multiphase microfluidics
Beschreibung:Date Completed 22.04.2015
Date Revised 17.06.2014
published: Print-Electronic
Citation Status MEDLINE
ISSN:1520-5827
DOI:10.1021/la5004484