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Electrochimical/Electrochemiluminescent Aptasensors Based on Tripropylamine Oxidation and Their Application in Protein Analysis

Author: LiuDongYuan
Tutor: YinXueBo
School: Nankai University
Course: Analytical Chemistry
Keywords: Aptamer Tripropylamine Electrochemiluminescence Modified electrode Electrochemical Ruthenium complexes
CLC: O629.73
Type: Master's thesis
Year: 2011
Downloads: 111
Quote: 0
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Abstract


The thesis is divided into three chapters, namely the introduction, the research and application of low potential electrochemiluminescence aptamer sensor, the TPA oxidation aptamer sensor protein analysis research and application. The introduction on the the aptamer definition, characteristics, development status summarized; discusses in detail the development of aptamer sensors, basic knowledge and application in the field of biochemistry; focuses on the principle of electrochemical sensors, development status and in the actual study. The second chapter is a low potential electrochemiluminescence aptamer sensor research and application. Electrochemiluminescence (ECL)-based biosensor is often used in the field of DNA and protein analysis. Although the electrochemiluminescence of the ruthenium complexes having a high sensitivity, but the high excitation potentials may lead to oxidative damage of biological molecules. This work was prepared as a lysozyme samples to be analyzed without damage, low potential electrochemiluminescence aptamer sensor. The single chain lysozyme aptamers first fixed to the surface of the gold electrode, and then combined to form a double-stranded DNA to its complementary DNA chain. Ru (phen) 32 electrochemiluminescence probe is inserted into the structure of double-stranded DNA. Stability of lysozyme and its aptamer binding leads to dissociation of double-stranded DNA, and the release of Ru (phen) 32. Can be observed in the double-stranded DNA modified electrode low potential electrochemical luminescence phenomenon, which is the double-stranded DNA can be pre-enrichment tripropylamine (TPA) and protonated TPAH proton release. While in the lysozyme aptamer as lysozyme special recognition component, a double-stranded DNA as the low potential electrochemiluminescence provide a microenvironment. The low potential electrochemiluminescence sensors, the detection limit of 0.45 pM lysozyme. Day accuracy (RSDs, n = 5) is less than 5%, the shows aptamers sensor reliability. Regeneration of the aptamer sensor verification of performance and low potential electrochemiluminescence can reduce oxidative damage of biological molecules. This method is successfully used for the direct analysis of diluted egg white sample. This work presents a promising high-sensitivity biological analysis platform, and can be applied to other low-potential ECL system development. Chapter the TPA oxidation aptamer sensor in protein analysis research and application. Propylamine (TPA) in the double-stranded DNA and single-stranded DNA-modified electrodes have different oxidation efficiency. Take advantage of this nature, the work of lysozyme, for example, the use of the TPA oxidation probe molecules to build a replacement electrochemical biosensor. A single-stranded DNA complementary to lysozyme aptamers gold sulfur key fixed to the surface of the gold electrode, and then double-stranded structure is formed on the electrode surface and Lysozyme aptamer binding. 10μL sample of lysozyme and its aptamer reaction, due to of lysozyme their aptamers high affinity, lysozyme replacement complementary strand, resulting in double-stranded DNA melting. Modified electrode in 20 mM TPA solution scan voltage 0.2-0.95V. The variation of the oxidation current is used for the quantitative content of lysozyme, the linear range of 1.0 pM to 1.1 nM, i.e. 6.0 × 106 lysozyme molecules can be detected. Preconcentration of TPA from the surface of the electrode due to the signal, the present method for high sensitivity at the same time, but also has a simple, stable, specific, time-saving, etc., and to avoid a complicated pretreatment of the sample and marker of the DNA chain. Direct analysis of the egg white samples to verify the feasibility of this biosensor, the recovery and reproducibility were 93.3-100% ,1.4-4 .2%.

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CLC: > Mathematical sciences and chemical > Chemistry > Organic Chemistry > Natural compounds > α-amino acids,peptides, proteins, nucleic acids > Protein
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