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Oriented immunomagnetic beads of zearalenone and preparation method and application of oriented immunomagnetic beads

A technology of zearalenone and immunomagnetic beads, which is applied in the preparation of test samples, material inspection products, measuring devices, etc., can solve the problem of affecting the antigen binding efficiency of directional immunomagnetic beads, the loss of antigen binding ability of antibodies, and the coupling Uncontrolled problems, saving time and cost, easy operation, and improved antibody binding ability

Inactive Publication Date: 2016-08-10
KONALD BIOTECH CO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The common directional immunomagnetic beads currently on the market are to directly couple the antibody to the surface of the magnetic bead, but due to the large size of the antibody molecule, the coupling is not controlled, and the antibody may be coupled to the magnetic bead in any direction
Certain antibody coupling directions will result in the inability to bind due to steric hindrance when the antigen molecule binds, and in some cases, the antigen-binding region of the antibody is coupled to the magnetic beads, causing the antigen-binding region to be blocked
Since the molecule of zearalenone is very small and carries only 1-2 antigenic epitopes, the blocking of an antigen-binding region may cause the antibody of this molecule to lose its antigen-binding ability
Therefore, it will affect the antigen binding efficiency of the whole directed immunomagnetic beads

Method used

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  • Oriented immunomagnetic beads of zearalenone and preparation method and application of oriented immunomagnetic beads
  • Oriented immunomagnetic beads of zearalenone and preparation method and application of oriented immunomagnetic beads
  • Oriented immunomagnetic beads of zearalenone and preparation method and application of oriented immunomagnetic beads

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1: Preparation of Zearalenone Directed Immunomagnetic Beads Using Gene Recombinant Protein G

Embodiment approach

[0043] A preferred embodiment of the present invention to prepare zearalenone-directed immunomagnetic beads is as follows:

[0044] 1. Magnetic bead activation

[0045] Take 5mg of carboxy-modified magnetic nano-beads, wash 3 times with 50mM PH6.0 MES buffer containing 0.01% SDS, and then suspend the magnetic beads with 5ml of 50mM PH6.0 MES buffer.

[0046] 2. Add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) 100mg to the magnetic beads, then add sulfur-N-hydroxysuccinimide (NHS) (Sulfo-NHS ) 100mg, react at room temperature for 15 minutes.

[0047] 3. After the reaction, the magnetic beads were washed 3 times with 50mM MES buffer pH6.0, and then resuspended in 5ml of 50mM MES buffer pH6.0.

[0048] 4. Add 20 mg protein G to the magnetic beads and react at room temperature for 4 hours.

[0049] 5. After the reaction, the magnetic beads were washed 3 times with 50mM MES buffer pH 6.0.

[0050] 6. After the reaction, the magnetic beads were blocked with 20ml of 1M eth...

Embodiment 2

[0058] Example 2: Preparation of Zearalenone Directed Immunomagnetic Beads Using Gene Recombinant Protein A

[0059] A preferred embodiment of the present invention to prepare zearalenone-directed immunomagnetic beads is as follows:

[0060] 1. Activation of magnetic beads.

[0061] 2. Take 5mg of carboxy-modified magnetic nano-beads, wash them 3 times with 50mM PH6.0 MES buffer containing 0.01% SDS, and then suspend the magnetic beads with 5ml of 50mM PH6.0 MES buffer.

[0062] 3. Add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) 100mg to the magnetic beads, then add sulfur-N-hydroxysuccinimide (NHS) (Sulfo-NHS ) 100mg, react at room temperature for 20 minutes.

[0063] 4. After the reaction, the magnetic beads were washed 3 times with 50mM MES buffer pH6.0, and then resuspended in 5ml of 50mM MES buffer pH6.0.

[0064] 5. Add 30 mg protein A to the magnetic beads and react at room temperature for 4 hours.

[0065] 6. After the reaction, the magnetic beads were wash...

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Abstract

The invention relates to oriented immunomagnetic beads of zearalenone and a preparation method and application of the oriented immunomagnetic beads. The oriented immunomagnetic beads are prepared by coupling protein G or protein A to magnetic beads, coupling a zearalenone antibody to the protein G or protein A located on magnetic beads; and utilizing a cross-linking agent to cross-link the protein G or protein A magnetic beads combined with the zearalenone antibody. According to the preparation method, the strong affinity between protein G / protein A and the antibody and the specific binding between protein G / protein A and an Fc segment of the antibody are fully utilized. The immunomagnetic beads are mainly used for binding and purifying zearalenone in grain, food, feed, milk, blood samples, and other multiple samples. The immunomagnetic beads can be used for pre-treatment of a sample before detection of zearalenone, and detection and purification of zearalenone in a sample.

Description

technical field [0001] The invention relates to directional immunomagnetic beads of zearalenone, a preparation method and application thereof, and belongs to the field of purification and detection of zearalenone. Background technique [0002] Zearalenone (ZEN), also known as F-2 toxin, is a 2,4-dihydroxybenzoic acid lactone compound with estrogen activity, and its chemical name is 6-(10-hydroxy-6- Oxycarbenyl)-β-resylate-μ-lactone. It is a mycotoxin produced by Fusarium graminearum and Fusarium yellow. It widely exists in nature and plays an important role in the growth and development of animals. It has a strong estrogen effect and can cause hyperestrogen syndrome in pigs, rats, mice, poultry and other animals and humans. It has reproductive and developmental toxicity, immunotoxicity, genotoxicity and suspected carcinogenicity. It mainly pollutes grain crops such as corn, wheat, and barley. Plcinta conducted a survey on the content of mycotoxins in grains and animal fe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/553G01N1/34
Inventor 张彦明柳家鹏
Owner KONALD BIOTECH CO