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Preparation and application of aflatoxin immunomagnetic beads

A technology of aflatoxin and magnetic absorption, applied in instruments, measuring devices, scientific instruments, etc., can solve the problems of a large number of organic reagents, time-consuming operation, environmental and operator damage, etc.

Pending Publication Date: 2022-05-06
SUZHOU KNOWLEDGE & BENEFIT TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, solid-phase extraction (SPE) columns are usually used to clean up and extract samples, but the operation of using SPE column to clean up and extract samples is time-consuming and requires a large amount of organic reagents, which is very harmful to the environment and operators.

Method used

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  • Preparation and application of aflatoxin immunomagnetic beads
  • Preparation and application of aflatoxin immunomagnetic beads

Examples

Experimental program
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Effect test

Embodiment 1

[0026] A preparation method for aflatoxin immunomagnetic microspheres, comprising the steps of taking magnetic microspheres with active groups on the surface, washing 1-5 times with 0.1M pH 8.4 borax buffer solution, adding 1 mg of magnetic microspheres per mg %-10% antibody ratio, add aflatoxin antibody, mix well at 25°C, react for 1 hour, after the reaction, wash the magnetic microspheres with borax buffer to wash away unreacted aflatoxin Antibody; add 10% BSA, mix well at 4-40°C, react for 1-5 hours, after the reaction, wash the magnetic microspheres with 50mM pH7.4 phosphate buffer to obtain aflatoxin immunomagnetic microspheres ball. The immunomagnetic microspheres were stored in buffer until use.

[0027] Its specific application is: pulverize calf feed with a pulverizer and pass it through a 5mm sampling sieve. Take 5g of the pulverized and sieved sample, add 20mL of 0.05M pH7.4 PBS buffer and mix well. Add 2 mg of aflatoxin immunomagnetic microspheres therein, and m...

Embodiment 2

[0029] A preparation method of aflatoxin immunomagnetic microspheres, comprising the steps of taking the magnetic microspheres with active groups on the surface, washing them twice with 0.05mM pH 6.0 citric acid buffer solution, and then dispersing the magnetic microspheres in the above-mentioned buffer in the liquid. According to the ratio of adding 1% antibody per mg of magnetic microspheres, add aflatoxin antibody, mix well at 4°C, and react for 17 hours. After the reaction, wash the magnetic microspheres with 0.05mM pH 6.0 citric acid buffer , to wash away unreacted aflatoxin antibodies; add 1mM ethanolamine solution, mix well at 37°C, and react for 1 hour. After the reaction, wash the magnetic microspheres with washing buffer to obtain aflatoxin immunomagnetic Microspheres. The immunomagnetic microspheres were stored in buffer until use.

[0030] Its specific application is: pulverize calf feed with a pulverizer and pass it through a 5mm sampling sieve. Take 5g of the ...

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Abstract

The preparation method comprises the following steps: washing magnetic microspheres with active groups on the surfaces by using a coupling buffer solution, adding an aflatoxin antibody to react with the magnetic microspheres, cleaning by using the coupling buffer solution after the reaction is finished, adding a blocking reagent to react, cleaning by using a cleaning buffer solution after the reaction is finished, and drying to obtain the aflatoxin immunomagnetic beads. The aflatoxin immunomagnetic microspheres are prepared. The surface of the magnetic microsphere is modified with an active functional group, so that the magnetic microsphere has reaction activity and can be quickly coupled with an aflatoxin antibody, and the magnetic microsphere has the capability of quickly capturing aflatoxin; when in use, a sample only needs to be crushed and then is fully and uniformly mixed with the immunomagnetic microspheres in a specific solution, the antibody on the surface of the magnetic microspheres can adsorb aflatoxin in the sample, and the aflatoxin sample with higher purity can be obtained by washing and eluting without a large amount of organic reagents.

Description

technical field [0001] The invention relates to the technical field of biological materials, in particular to the preparation and application of aflatoxin immune magnetic beads. Background technique [0002] Aflatoxin (AF), a difuran ring toxoid produced by certain strains of Aspergillus flavus and Aspergillus parasiticus. There are about 20 derivatives, among which aflatoxin B1 is the most toxic and carcinogenic. After animals eat aflatoxin-contaminated feed, trace amounts of toxins can be detected in liver, kidney, muscle, blood, milk, and eggs. When humans eat soybeans, corn, sorghum, etc. contaminated with aflatoxin or contaminated animal products, aflatoxin will accumulate in the human body and eventually lead to poisoning. In 1993, aflatoxin was classified as a class of naturally occurring carcinogens by the Cancer Research Institute of the World Health Organization (WTO), which is a highly toxic and highly toxic substance. [0003] At present, the main methods for ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543
CPCG01N33/54326
Inventor 阳承利李雪
Owner SUZHOU KNOWLEDGE & BENEFIT TECH CO LTD
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