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Novel Methods of Electrochemical Biosensing Based on Enzyme Amplification

Author: WangCui
Tutor: YuRuQin
School: Hunan University
Course: Analytical Chemistry
Keywords: Electrochemical biosensor Electronic channel switching-basedaptasensor Circular strand-displacement polymerase reaction Hybridization chain reaction Streptavidin-alkaline phosphatase DNA IgE ATP
CLC: O657.1
Type: Master's thesis
Year: 2013
Downloads: 24
Quote: 0
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Abstract


Due to the advantages of simple operation, high sensitivity, low cost, fastresponse, electrochemical sensors have been widely used for trace analysis. This thesisused the irreversible oxidation peak as the electrochemical signal which produced byalkaline phosphatase converting electrochemically inactive1-naphthyl phosphate to anelectrochemically active1-naphthol. By combining the DNA modification techniques,hairpin probes, amplification techniques, this research paper develops several novelelectrochemical biosensors based on the traditional method. This thesis achieveseffective detection of DNA, protein and small molecules and completes three tasks.The detailed content is described as follows:（1） By combing the circular strand-displacement polymerase reaction （CSDPR）and the hybridization chain reaction （HCR）, we developed a novel electrochemicalstrategy for ultrasensitive DNA detection. In this assay, hybridization of hairpin-shaped capture DNA to target DNA resulted in a conformational change of the captureDNA with a concomitant exposure of its stem. The primer was then hybridized withthe exposed stem and triggered CSDPR. Furthermore, the free part of the primerpropagated a chain reaction of hybridization events between two DNA hairpin probeswith biotin labels, enabling an electrochemical reading using the streptavidin-alkalinephosphatase. The proposed biosensor showed to have very high sensitivity andselectivity with a dynamic response range covering through10fM to1nM, and thedetect limit was as low as8fM. This strategy could also be applied in complexbiological systems. This strategy paved a way for molecular diagnostics in complexbiological systems.（Chapter2）（2） Based on the combination of two biomaterials （one aptamer and one enzyme）,an electronic channel switching-based aptasensor for ultrasensitive IgE detection isdeveloped. This strategy proposes an innovative sensing concept: the surface-immobilized IgE serve as a barrier that separates enzyme from its substrate. Thisaptasensor exhibits a low detection limit of4.44×10-6μg mL-1and a wide dynamicresponse range from4.44×10-6to4.44μg mL-1. More importantly, this electronicchannel switching-based sensor may open a new avenue for the detection of proteinsand other biomacromolecules.（Chapter3）（3） By introducing the two fragments of ATP aptamer and HCR amplification, we developed a novel electrochemical sandwich assay for ATP detection. In this assay, wedesign a link DNA which includes the sequence of one fragment of ATP aptamer andthe trigger sequence for HCR. Therefore, this proposed biosensor effectivelyintroduced HCR to the sensor, achieved a novel electrochemical biosensor based onsandwich structure and HCR amplification. The results show that this method has theadvantages of low background, high selectivity and simple operation.（Chapter4）

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CLC: > Mathematical sciences and chemical > Chemistry > Analytical Chemistry > Instrument analysis ( physics and physical chemistry ) > Electrochemical analysis
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