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Microcystin -LR type immune electrochemical impedance detection method

Author: ShiHui
Tutor: WangHongXin
School: Jiangnan University
Course: Nutrition and Food Hygiene
Keywords: Microcystin -LR Polyclonal antibody Immunosensor AC impedance spectroscopy Room temperature ionic liquids
CLC: R123.1
Type: Master's thesis
Year: 2009
Downloads: 144
Quote: 1
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Abstract

Microcystin-LR (MC-LR) is the most common freshwater bodies and the most toxic of a class of cyanobacterial toxins. At present, the algal toxin detection methods require expensive equipment and the operation more complicated and can not meet the rapid detection. Therefore, the study and the establishment of a rapid, sensitive and simple detection method has great significance. In this paper, MC-LR completely synthetic antigen polyclonal antibody preparation specific basis, combined immune sensing technology and nanomaterials self-assembly method to build electrochemical impedance type immune sensor detection methods, the main contents are as follows: The \active ester method \New Zealand white rabbits immunized polyclonal antibodies obtained after purification its potency is higher than 5.2 × 10 ^{4 . By indirect competitive ELISA method, MC-LR competition standard curve linear range of 0.25 ~ 4μg / L; half inhibition rate IC50 was 0.68μg / L. In addition to the intersection with the MC-RR response rate of about 21.9%, but with the MC-LW, MC-LF cross-reactivity were less than 1.0%. Based on L-cysteine ??(L-Cys) and gold nanoparticles (Nano-Au) fixed antibody technology to build a MC-LR unmarked type immune sensor detection method. The impedance for MC-LR linear response range of 0.05 ~ 300 ng / mL; minimum detection limit of 1.82 × 10 ^{-2 ng / mL (S / N = 3); detection time of only 15 min; source of drinking water for the average recoveries were between 96.8% ~ 112.5%; modified electrode cyclic scan on lap 42, the peak current is no significant change in coefficient of variation was 3.58%; 4 ℃ for 30 days under the Stability Good. Based on 1,6 - hexanedithiol (HDT) self-assembly technology and nano gold adsorption method of combining fixed microcystin antibody construct immune sensors. The results show that the linear response range of the impedance 0.25 ~ 95 ng / mL, the detection limit was 8.5 × 10-2 ng / mL (S / N = 3); electrode reproducibility and stability, for the same density continuous Determination of eight times, and the relative standard deviation was 7.84%; 4 ℃ after 30 days of storage no significant change in the impedance response signal; applied to drinking water, the sample average recovery was 92.6% ~ 118.3%; detection time is 15 min. Based on multi-walled carbon nanotubes (MWNTs) / room temperature ionic liquids (RTILs) modified electrode fixed antibody constructed MC-LR Immunosensors. Act with a higher sensitivity, detection limit was 1.7 × 10 ^{-3 ng / mL (S / N = 3), at 5 × 10-3 ~ 1 × 10 ^{2 ng / mL range a good linear correlation; drinking water average recoveries were 80.95% ~ 127.23%; common ions in water for determination of MC-LR was no significant interference. Protective solution was first proposed electrode design concept. Based on the excellent characteristics of ionic liquids, at 4 ℃, for the stability of the modified electrodes up to 60 days of investigation, screened out of favor with antibody protein to maintain its stability and its optimal concentration of ionic liquids. The use of circular dichroism initially explored RTILs to maintain the stability of the mechanism of antibody proteins, revealing the antibody protein conformational changes in RTILs. The developed detection of MC-LR in drinking water impedance type electrochemical immunoassay sensing detection methods, sensitive, rapid, convenient and has good development prospects for the realization of direct, real-time, in-situ, on-line MC-LR daily monitoring provides important experimental basis.}}}}

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