Bioluminescent Protein-Inhibitor Pair in the Design of a Molecular Aptamer Beacon Biosensing System

Angeliki Moutsiopoulou, David Broyles, Hamdi Joda, Emre Dikici, Avinash Kaur, Angel Kaifer, Sylvia Daunert, Sapna K. Deo

Research output: Contribution to journalArticle

Abstract

Although bioluminescent molecular beacons designed around resonance quenchers have shown higher signal-to-noise ratios and increased sensitivity compared with fluorescent beacon systems, bioluminescence quenching is still comparatively inefficient. A more elegant solution to inefficient quenching can be realized by designing a competitive inhibitor that is structurally very similar to the native substrate, resulting in essentially complete substrate exclusion. In this work, we designed a conjugated anti-interferon-γ(IFN-γ) molecular aptamer beacon (MAB) attached to a bioluminescent protein, Gaussia luciferase (GLuc), and an inhibitor molecule with a similar structure to the native substrate coelenterazine. To prove that a MAB can be more sensitive and have a better signal-to-noise ratio, a bioluminescence-based assay was developed against IFN-γand provided an optimized, physiologically relevant detection limit of 1.0 nM. We believe that this inhibitor approach may provide a simple alternative strategy to standard resonance quenching in the development of high-performance molecular beacon-based biosensing systems.

Original languageEnglish (US)
Pages (from-to)7393-7398
Number of pages6
JournalAnalytical Chemistry
Volume92
Issue number11
DOIs
StatePublished - Jun 2 2020

ASJC Scopus subject areas

  • Analytical Chemistry

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