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Study on the Colorimetric Detection for Thrombin Protein Based on G-quadruplex DNAzyme and Nano-aptasensors

Author: CuiJing
Tutor: ChengGuiFang；HePinGang；FangYuZhi
School: East China Normal University
Course: Analytical Chemistry
Keywords: Gold nanoparticles DNA enzyme Magnetic nanoparticles Aptamer Thrombin Colorimetric
CLC: TP212.3
Type: Master's thesis
Year: 2011
Downloads: 126
Quote: 1
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Abstract


The Human Genome Project (HGP) the successful completion of a declaration of the post-genomic era, life science research shifted from gene sequencing to functional genomics and proteomics research up. Proteins are the basis of living matter, throughout the life activities of all processes, and human health are closely related. Study of protein structure and function in human nutrition and health research, drug screening, diagnosis and clinical analysis and other aspects are important. How to build a high-throughput, high sensitivity, high specificity for protein detection technology to become the current proteomics research hotspot. Quantitative detection of the protein based on the study of proteins. Traditional protein detection mainly by antigen - antibody interactions. The aptamer (Aptamer) emergence up deficiencies of existing antibodies, are widely used in biological sensor specific detection of the protein. G-tetrad DNA enzyme as a novel DNA enzyme with peroxide catalytic activity, easy to build and can be used with the ECL biosensor and colorimetric sensors, to achieve the target molecule signal amplification detection. In addition, nanoparticles synthesized simple, low cost, large specific surface area, good biocompatibility and optical, electrical properties, the biosensor has been widely used. Thus, the development of new nano-biosensor protein is expected to achieve rapid, highly sensitive detection. The innovation of this paper is to build a G-tetrad based on the new nano DNA enzyme colorimetric biosensor. Combined with excellent biocompatibility gold nanoparticles and magnetic nanoparticles paramagnetic developed a new specific recognition protein sensing technology, pioneering the post-genome era of research. The main contents are as follows: The first chapter This chapter first introduces the principle and classification of biosensors, which focuses on the classification and Biosensor Research. Detailed overview of G-tetrad DNA enzyme structure, nature and its DNA and protein detection, and introduced nanomaterials in biosensor applications, illustrates the theoretical basis of this thesis and topics in accordance. The thesis concludes with the purpose and significance. The second chapter is based on double gold DNA enzyme catalysis and separation of magnetic nanoparticles highly sensitive colorimetric detection of thrombin protein research paper based on gold nanoparticles and DNA enzyme catalyzed double magnetic nanoparticles to thrombin (thrombin) for the study Construction of a new colorimetric nanosensors. Gold nanoparticles surface markers on a kind of catalytic properties of enzymes and DNA aptamer (TBA), and the fixed magnetic nanoparticles in a complementary nucleic acid hybridization to form aptamer sensors. In the presence of thrombin, tend aptamer binding with thrombin to form G-tetrad structure, making the original double-stranded dissociation enzyme labeled DNA liberated gold particles. Free out of the DNA enzyme labeled gold particles have a dual function catalyst in the presence of H2O2, can catalyze the oxidation of ABTS oxidation products generated blue-green, to the amplification of thrombin colorimetric detection purposes. Because of the high sensitivity using two catalyst, the sensor can detect the above thrombin 0.1nM. This detection method is low cost, simple, rapid, high sensitivity, specificity and other biological molecules is expected to provide a variety of means testing. The third chapter is based on symmetric multi-stage build DNA enzyme biosensor sensitivity colorimetric detection of thrombin proteins symmetric multi-enzyme is a DNA compared to normal DNA enzyme can be a style more sensitive detection of nucleic acid or protein G-quadruplex sub-structure of the new activity of the enzyme, thus widely used in a variety of biomolecules detection on. This combination of symmetric multi-enzyme and DNA aptamer technology to thrombin (thrombin) designed for the study of two sensitive colorimetric signal is amplified aptamer sensor. \It uses the aptamer (TBA) high specificity for thrombin recognition and multi-symmetric signal components to amplify DNA enzyme to build, can be used for detection of thrombin. The aptamer sensor is simple, low cost, but susceptible to external interference detection of false positive signals, affecting target detection. \The thrombin aptamer sensor detects a concentration of 1 to 40nM thrombin has a good linear relationship between the detection limit of 1nM. This new type of nano-biosensors making simple, detecting a high specificity and sensitivity, and is expected to be applied in the biomedical field detection of thrombin.

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