Synthesis and sensing integration: A novel enzymatic reaction modulated Nanoclusters Beacon (NCB) “Illumination” strategy for label-free biosensing and logic gate operation

A novel fluorescent label-free “turn-on” NAD+ and adenosine triphosphate (ATP) biosensing strategy is proposed by fully exploiting ligation triggered Nanocluster Beacon (NCB). In the presence of the target, the split NCB was brought to intact, which brought the C-rich sequence and enhancer sequence...

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Bibliographic Details
Published in:	Biosensors & bioelectronics 2016-12, Vol.86, p.588-594
Main Authors:	Hong, Lu, Zhou, Fu, Wang, Guangfeng, Zhang, Xiaojun
Format:	Article
Language:	eng
Subjects:	Adenosine triphosphate Adenosine Triphosphate - analysis Ag nanocluster Beacons Biosensors Computers, Molecular Detection DNA - chemistry DNA - radiation effects Equipment Design Equipment Failure Analysis Feasibility Studies Fluorescence Label-free Lighting - instrumentation Logic circuits Logic gate Microscopy, Fluorescence - instrumentation NAD NAD - analysis Nanoclusters Beacon (NCB) Nanoconjugates - chemistry Nanoconjugates - radiation effects Nanostructure Reproducibility of Results Sensitivity and Specificity Signal Processing, Computer-Assisted - instrumentation Silver - chemistry Staining and Labeling Strategy
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Summary:	A novel fluorescent label-free “turn-on” NAD+ and adenosine triphosphate (ATP) biosensing strategy is proposed by fully exploiting ligation triggered Nanocluster Beacon (NCB). In the presence of the target, the split NCB was brought to intact, which brought the C-rich sequence and enhancer sequence in close proximity resulting in the lightening of dark DNA/AgNCs (“On” mode). Further application was presented for logic gate operation and aptasensor construction. The feasibility was investigated by Ultraviolet–visible spectroscopy (UV–vis), Fluorescence, lifetime and High Resolution Transmission Electron Microscopy (HRTEM) etc. The strategy displayed good performance in the detection of NAD+ and ATP, with the detection limit of 0.002nM and 0.001mM, the linear range of 10–1000nM and 0.003–0.01mM, respectively. Due to the DNA/AgNCs as fluorescence reporter, the completely label-free fluorescent strategy boasts the features of simplicity and low cost, and showing little reliance on the sensing environment. Meanwhile, the regulation by overhang G-rich sequence not relying on Förster energy transfer quenching manifests the high signal-to-background ratios (S/B ratios). This method not only provided a simple, economical and reliable fluorescent NAD+ assay but also explored a flexible G-rich sequence regulated NCB probe for the fluorescent biosensors. Furthermore, this sensing mode was expanded to the application of a logic gate design, which exhibited a high performance for not only versatile biosensors construction but also for molecular computing application. •A simple label-free biosensor was developed for NAD+ detection with DNA/AgNCs probe.•The specific NAD+-ligation triggered nanocluster beacon from split to intact.•Logic gate operation was investigated based on the NAD+ sensor.•The construction of ATP aptasensor showed the platform universal.•The platform is highly sensitive and selective.
ISSN:	0956-5663 1873-4235