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Investigation of Assay of Glucose and MicRNA via Colorimetric Biosensors

Author: ZhangYanJun
Tutor: WangJianXiu
School: Central South University
Course: Analytical Chemistry
Keywords: Glucose MicRNA Colorimetry Biosensors
CLC: O657.3
Type: Master's thesis
Year: 2013
Downloads: 20
Quote: 0
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Abstract


Since the development of the first-generation glucose biosensors, biosensing techniques have been gradually fused into different disciplines, such as chemistry, biology, and medicine. Optical biosensors have been constructed via examining the absorption, fluorescence, scattering, chemiluminiscence, and other optical properties, which are closely related to the chemical information of the analytes determined. Due to their high sensitivity and selectivity, rapidness, and accurateness, optical biosensors have been widely used in many fields. In this thesis, relying on the specific properties of the nanomaterials and enzymes, two colorimetric biosensors for assay of glucose and micRNA have been constructed.Recently, the use of biomaterials as reductants for the synthesis of nanoparticles is a novel area in nanobiotechnology. The change in the chemical and physical properties of the as-formed nanoparticles forms the basis for the design of various biosensors. In this thesis, one step synthesis of Ag NPs using H2O2as a reductant under UV irradiation was reported. The Ag NPs was biocatalytic grown without adding any Ag NP seeds, and the whole procedure was simple, fast and efficient. The Ag nanoparticles exhibit yellow color, which could be clearly seen by the naked eyes. The UV-vis absorption of the Ag NPs is dependent on the concentrations of H2O2, thus a facile colorimetric method for assay of H2O2has been established. Via integrating the catalytic activity of glucose oxidase with the enhanced absorption signals of the Ag NPs, successful determination of glucose in human blood samples has been proposed.MicroRNA (micRNA) belongs to a family of small non-protein coding RNAs (typically17-25nucleotides long). The expression level of micRNA could provide useful information for clinical diagnosis and prognosis of a large variety of human cancers. In this thesis, a colorimetric biosensor for assay of micRNA has been fabricated via the catalytic reaction of the poly-enzyme. A biotinylated micRNA (biotin-micRNA) whose sequence is the same as that of a micRNA target is allowed to compete with the micRNA target for the oligonucleotide (ODN) probe preimmobilized onto the wells of a96-welll plate. This step was followed by the complexation of the biotin-micRNA with (Ploy-HRP)/streptavidin conjugates. The poly-HRP could effectively catalyze the H2O2-mediated oxidation of3,3’,5,5’-tetramethylbenzidine (TMB) with the formation of the colored products. Due to the competitive hybridization reaction, the increase in the concentration of target micRNA leads to the decrease in the UV-vis absorption signals. The method is simple, highly reproducible, and selective, being of great significance for early diagnosis of glioma patients.

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