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The Study of Label-Free Impedimetric Immunosensor for AFB1

Author: WuNingNing
Tutor: CaoLiXin
School: Harbin Institute of Technology
Course: Chemical Engineering and Technology
Keywords: Aflatoxin B1 Unmarked impedance immunosensor Poly -o-phenylenediamine modified electrode Electrochemical impedance spectroscopy
CLC: TP212
Type: Master's thesis
Year: 2011
Downloads: 29
Quote: 0
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Abstract


This article provides an overview of the immune sensor and the research progress and application of the modified electrode to analyze the advantages and disadvantages of AFB1 detection method is to study the preparation conditions of the \and electrochemical impedance spectroscopy (EIS) on AFB1 quantitative detection achieved good detection effect. First, the use of the modified electrode prepared by cyclic voltammetry PoPD, studies show that in 0.05 mol / L o-PD 1 mol / L H2SO4 electro-polymerization solution, -600 mV 1000 mV potential range to 50 mV / s scan rate, a cycle of 30 weeks can get to contain a large number of amino-NH2 \The film not only having a good penetration of the select characteristics, can make H choose to penetrate into the membrane and a catalytic reduction reaction, and the oxidation of H / H. Then, based PoPD modified electrode, the fixed conditions of the preparation conditions and AFB1 AFB1 immunosensor antigen. The results showed that at 25 ℃ glutaraldehyde crosslinked, anti-AFB1 purified antibody fixed closed and fetal calf serum, and the operation time of the three should be for 45 min; 80% PBS 20% methanol as the reaction solution, pH = 7.40, combined with a temperature of 30 ° C can be obtained the AFB1 antigen fixed amount. Secondly, the reaction occurring in the assembling process of the AFB1 immune sensor. Glutaraldehyde PoPD / Au electrode crosslinking and antibody fixed, the two aldehyde groups of glutaraldehyde-CHO respectively PoPD amino-NH 2 and the residual amino groups of the antibody immunoglobulin - NH 2 occurred Schiff base reaction, formation of a stable Ar-N = C (CH 2 ) 3 C = NR structure, thereby the antibody is fixed on the electrode surface, and is formed with molecular recognition function of AFB1 immunosensor. Again, the EIS method, electrochemical characterization of the the AFB1 immune sensor assembly process and build the model of the molecular recognition membrane interface. Popd / Au electrode surface with glutaraldehyde cross-linked, closed performed anti-AFB1-antibody fixed and calf serum, electron transfer resistance Drill increased; performing AFB1 antigen-specific immune response, the electron transfer resistance increases with AFB1 immune binding capacity increases; H, non-specific adsorption, and the unevenness of the surface of the electrode, AFB1-immune sensor EIS spectra has obvious \Finally, the use of the AFB1 immune sensors, EIS method of AFB1 detection. The results show that having a two-stage linear relationship 0.03μg/mL 0.7μg/ml range. When the concentration range of 0.03 and 0.1μg/mL, linear equations ΔRP =-5960Ω 2.75 × 10 11 × cAFB1, R2 = 0.9946; When the concentration range for 0.1μg/mL 0.7μg/mL time, linear equations ΔRP = 19410Ω 2.49 × 1010 × cAFB1, R2 = 0.9807; lowest limit of detection LOD of 2.75 × 10 -8 g / mL. Further, the solution of AFB1 the immunosensor immune sensor chaotropic and preservation methods were optimized. Studies have shown that in 0.2 mol / L glycine, 0.2 mol / L HCl (pH = 2.60) in the dissociation can be reused 10 times. Wet save the AFB1 immunosensor temporarily save and dry the longest shelf life of 11 days. Compared with the same types of unmarked impedance-type the AFB1 immune sensor, the method has good linearity, the lowest detection limit of 1 × 10-7 g / mL LOD less than reported in the literature. The immunosensor can be reused, the preparation time is short, low cost, has great commercial potential and market value.

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