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Study of Novel Bio-analytical Methods Based on Biological and Chemical Reactions

Author: LiYun
Tutor: LiGenXi
School: Nanjing University
Course: Biochemistry and Molecular Biology
Keywords: Nitrite Platinum nanoparticles Griess reaction Chemically modified electrode Electrochemical End protection Nicking endonuclease Interaction Folic acid Folate receptor
CLC: Q503
Type: Master's thesis
Year: 2011
Downloads: 62
Quote: 0
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Abstract


Biochemical analysis of development plays a very important role in solving human health problems and environmental crisis. There are many biological and chemical reactions in nature, with the development of these reactions biochemical analysis has been a hot topic in academia. Therefore, we can take advantage of a number of organic reactions and new methods of research carried out biochemical analysis based on the specific reaction of biological affinity, specific contents include: 1. Study of the new method based the Griess the reaction development nitrite Electrochemical detection of nitrite can cause infant methemoglobinemia and hemoglobin binding, and related to the formation of carcinogenic nitrosamines class. Therefore, we designed a way to sensitive and simple detection method of drinking water nitrite concentration is increasingly important for the human health and environmental health. In this chapter, we propose a novel electrochemical biosensor for detection of nitrite functionalized platinum nanoparticles (PtNPs). First, the surface of the gold electrode modified 4 - (2 - aminoethyl) aniline, and then modified in the surface of the platinum nanoparticles 5 - [1, 2] dithiolan-3-yl-PENTANOIC acid [2 - (naphthalene-1-ylamino) -ethyl] amide (DPAN). When the presence of nitrite ion in the solution, the surface of a gold electrode modified 4 - (2 - aminoethyl) aniline will the PtNPs surface of DPAN Griess reaction. Thereby, PtNPs connected to the surface of the gold electrode, and generated by H2O2 reduction reaction the use PtNPs electrocatalytic electrochemical signal can achieve the purpose of the detection of nitrite ion. The method detects the linear range of the nitrite ion concentration is 10-1000μM, detection limit reached 5μM. In the experiment, we successfully used this method to detect the nitrite ion in the lake, the test results and standardized detection of nitrite ion UV - visible spectrum detection method. It can be seen, this method has the potential value of the nitrite concentration in the monitoring of drinking water daily. Based endonuclease nicked auxiliary amplification reaction electrochemical analysis of the interaction of proteins with small molecules combined with moderate affinity and specificity of small organic molecules of a particular protein can not only interfere with protein function is used for chemical genetic studies, and can locate the protein and for protein quantitative molecular biology diagnostic, test new methods of interaction between small molecules and proteins will contribute to the development of molecular biology diagnostic and treatment technology. In this chapter, we combine end protection and nicking endonuclease assisted amplification reaction (NEA), folate receptor - the interaction between folic acid, for example, proposed a new electrochemical method to detect protein - small molecule role. First of all, the end to protect the specific binding of folate on folate receptor ssDNA molecules, folic acid - folic acid receptor complex can protect ssDNA from the hydrolysis of exonuclease I (Exo I). Secondly, NEA reaction i.e. nicking enzyme can identify double-stranded DNA and only the specific nucleotide sequence wherein a cutting chain. Due to the release of the other strand can be immobilized on the electrode surface with a specific sequence of nucleotide hybridization, in order to initiate the second round of the cleavage reaction, after multiple cycles, the surface of the gold electrode a nucleic acid probe the electric signal molecules in the solution [ Fe (CN) 6] 3 - / 4 impediment is gradually reduced, so that the electrochemical signal increases, reaches the sensitive detection of proteins - The purpose of the interaction between small molecules. Compared to other protein - small molecule interactions detection method, this method has the advantages of high sensitivity, high selectivity, high-volume, low-cost and easy to operate, and provide a new future detection of protein - small molecule interactions ideas.

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