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Magnetic multi-sensibilization sensor for aflatoxin as well as preparation and application of magnetic multi-sensibilization sensor

A technology of aflatoxin and sensor, which is applied in the field of grain, oil and food safety, can solve problems such as fastness, convenience, low cost, long time-consuming instrument detection methods, unfavorable on-site operation and use, etc., and achieves high adaptability to extreme conditions and a preparation method Mature and stable, weak toxicity effect

Active Publication Date: 2021-08-06
JIANGSU UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the instrument detection method is time-consuming, complicated to operate, expensive equipment, and requires professional personnel, which is not conducive to on-site operation and use, and cannot meet the requirements of fast, convenient, and low-cost detection.
Immunological detection methods include ELISA, colloidal gold test strips, and immunoaffinity columns, which are mainly used for the screening and detection of AFB1 in a large number of samples. They are fast, cheap, low-cost and specific, and are very suitable for a large number of samples. However, there is a certain gap between the detection sensitivity and the instrument detection method

Method used

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  • Magnetic multi-sensibilization sensor for aflatoxin as well as preparation and application of magnetic multi-sensibilization sensor
  • Magnetic multi-sensibilization sensor for aflatoxin as well as preparation and application of magnetic multi-sensibilization sensor
  • Magnetic multi-sensibilization sensor for aflatoxin as well as preparation and application of magnetic multi-sensibilization sensor

Examples

Experimental program
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Effect test

preparation example Construction

[0041] (1) His-Fe 3 o 4 Preparation of nanozymes

[0042] FeCl 3 ·6H 2 O was dissolved in ethylene glycol solution, stirred until the solution was transparent, and CH was added 3 COONa and histidine were stirred at 800-1200rpm. After 30min, the mixture was ultrasonicated for 10min, and then transferred to a polytetrafluoroethylene-lined stainless steel autoclave at 200-240°C for 12-16h. After the reaction was completed, the high-pressure The kettle was cooled to room temperature, washed several times with ethanol and dried at 60°C to obtain His-Fe nanoparticles with a size of 400-500 nm. 3 o 4 , namely His-Fe 3 o 4 nanozyme;

[0043] (2) Capture probe His-Fe 3 o 4 Preparation of @Ab

[0044] Antibodies and His-Fe 3 o 4 The histidine on the surface of the nanozyme is coupled by the activated ester method, the monoclonal antibody used is dissolved in phosphate buffer solution, and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide salt is added acid salt (EDC) and N-hyd...

Embodiment 1

[0060] A kind of preparation of aflatoxin magnetic multiple sensitization sensor, comprises the following steps:

[0061] 1) His-Fe 3 o 4 Preparation of nanozymes:

[0062] 0.27g FeCl 3 ·6H 2 O was dissolved in 10 mL of ethylene glycol solution, and after the solution was clarified, 1.8 g of CH was added 3 Mix COONa and 0.5g histidine, keep stirring for 30min, then sonicate for 10min, react in a polytetrafluoroethylene-lined stainless steel autoclave at 200°C for 16 hours at high temperature, cool to room temperature, wash with ethanol 3 to 5 times, and dry at 60°C 12h, His-Fe with a size of 400-500nm can be synthesized 3 o 4 nanozyme;

[0063] 2) Capture probe His-Fe 3 o 4 @Ab's preparation:

[0064] Dissolve the monoclonal antibody used in phosphate buffer solution, add EDC and NHS for activation reaction, the ratio of the three is 1:40:40, respectively 28.3μmol, 1.132mmol and 1.132mmol, incubate at room temperature for 30min, ultrafilter Remove unreacted material...

Embodiment 2

[0070] Based on the application of the sensor prepared in Example 1 on aflatoxin B1 (AFB1) in food, the specific detection method is as follows:

[0071] There are seven groups of AFB1 standard solutions with different concentrations, the volume is 1.5mL, the concentrations are 0, 0.0001, 0.0005, 0.001, 0.005, 0.01, 0.05, 0.1, 0.5, 1ng / mL, add 0.4mL of capture probe, 25℃ Incubate at constant temperature and shake for 15 minutes, remove the supernatant of the complex constructed by magnetic separation, add 0.4 mL of detection probe, and incubate at 25°C for 15 minutes with constant temperature and shake, remove the supernatant to construct the spatial structure of magnetic separation, add 0.4 mL of chromogenic reagent, Incubate at 25°C with constant temperature and shaking for 5 minutes, magnetically separate and precipitate the spatial structure, measure the UV absorbance of the supernatant, and establish a standard curve between the absorbance and AFB1;

[0072] The prepared ...

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Abstract

The invention discloses a magnetic multi-sensibilization sensor for aflatoxin as well as preparation and application of the magnetic multi-sensibilization sensor. The nano enzyme (A) is formed by modifying ferroferric oxide with histidine; the capture probe (B) is formed by coupling a nano-enzyme with a monoclonal antibody of aflatoxin B1 (AFB1); the detection probe (C) is formed by coupling a nano-enzyme with an antigen of AFB1; the capture probe and the detection probe form a two-dimensional net structure (D) through specific binding between antigens and antibodies. The method comprises the following steps: preparing magnetic nano-enzyme for later use; modifying the capture probe; modifying the detection probe; and forming a two-dimensional net structure. In the detection process, after the constructed two-dimensional network structure is subjected to magnetic separation and enrichment, a 3, 3 ', 5, 5'-tetramethyl benzidine (TMB) color developing reagent is added, and the two-dimensional network structure catalyzes the substrate to develop color. The magnetic multi-sensibilization detection method established by the invention has the advantages of high enrichment degree, high sensitivity, rapidness and the like, and is suitable for large-scale screening detection of AFB1 in food.

Description

technical field [0001] The invention relates to a nano-enzyme catalyzed ultraviolet signal detection technology, which is suitable for the fields of grain, oil, food safety and the like, and in particular relates to a magnetic multi-sensitization sensor for aflatoxin and its preparation and application. Background technique [0002] Aflatoxins are a class of compounds with similar chemical structures. The main toxin-producing fungi include Aspergillus flavus and Aspergillus parasiticus. Among the known mycotoxins, aflatoxin is the most toxic and most harmful to human health. It is one of the most toxic substances in the world. Among them, the most toxic and widely distributed type of aflatoxin is Aspergillus flavus Toxin B1 (AFB1). Due to its strong carcinogenic, teratogenic and mutagenic effects, AFB1 was classified as the first class of carcinogens by the International Agency of Research on Cancer (IARC) in 1988. [0003] Aflatoxin B1 can cause extensive pollution to agr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78G01N21/33G01N33/577
CPCG01N21/78G01N21/33G01N33/577
Inventor 孔德昭鞠嘉和卢勇莉唐盛沈薇
Owner JIANGSU UNIV OF SCI & TECH
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