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A kind of a-type trichothecenes toxin polyclonal antibody and its preparation method and application

A trichothecenes, polyclonal antibody technology, applied in the direction of antifungal/algae/lichen immunoglobulin, animal/human protein, serum albumin, etc., can solve the problems of limited scope of use and complicated operation, Achieve good specificity and sensitivity, high similarity, and good immunogenicity

Active Publication Date: 2018-04-24
GUANGDONG OCEAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method uses gas phase as the main instrument for detection. The sample needs to be derivatized before detection, and the operation is complicated. It can only detect two types of trichothecenes in traditional Chinese medicine, and the scope of application is relatively limited.

Method used

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  • A kind of a-type trichothecenes toxin polyclonal antibody and its preparation method and application
  • A kind of a-type trichothecenes toxin polyclonal antibody and its preparation method and application
  • A kind of a-type trichothecenes toxin polyclonal antibody and its preparation method and application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] The preparation method of type A trichothecene toxin artificial antigen and antibody is prepared by the following steps:

[0051] (1) Design and synthesis of hapten:

[0052] In order to prepare anti-type A trichothecene polyclonal antibody and coating agent, synthesize the group hapten 3-Ac-NEOS of type A trichothecene; the synthesis of 3-Ac-NEOS:

[0053] 1) Using T-2 toxin as the reaction substrate, add anhydrous pyridine and acetic anhydride to acetylate the C3 hydroxyl group of T-2 toxin to prepare 3-acetyl-T-2, and pass the reaction product through silica gel column For purification, use n-hexane / ethyl acetate 50:45 as eluent. After the eluate was concentrated, it was identified by mass spectrometry;

[0054] Mass Spectrometry: The m / z of the strongest ion peak detected is 524.66[M+18] + , the molecular weight obtained is 508, which is consistent with the theoretical molecular weight of the target product 3-Ac-T-2 toxin;

[0055] 2) Treat 3-acetyl-T-2 with rat...

Embodiment 2

[0074] The preparation method of type A trichothecene toxin artificial antigen and antibody is prepared by the following steps:

[0075] (1) Design and synthesis of hapten:

[0076] In order to prepare type A trichothecene toxin polyclonal antibody and coating agent, synthesize the group hapten 3-Ac-NEOS of type A trichothecene toxin; the synthesis of 3-Ac-NEOS:

[0077] 1) Using T-2 toxin as the reaction substrate, add anhydrous pyridine and acetic anhydride to acetylate the C3 hydroxyl group of T-2 toxin to prepare 3-acetyl-T-2, and pass the reaction product through silica gel column For purification, use n-hexane / ethyl acetate 50:50 as eluent. After the eluate was concentrated, it was identified by mass spectrometry;

[0078] Mass Spectrometry: The m / z of the strongest ion peak detected is 526.66[M+18] + , the molecular weight obtained is 508, which is consistent with the theoretical molecular weight of the target product 3-Ac-T-2 toxin;

[0079]2) Treat 3-acetyl-T-2 wi...

Embodiment 3

[0098] The preparation method of type A trichothecene toxin artificial antigen and antibody is prepared by the following steps:

[0099] (1) Design and synthesis of hapten:

[0100] In order to prepare type A trichothecene toxin polyclonal antibody and coating agent, synthesize the group hapten 3-Ac-NEOS of type A trichothecene toxin; the synthesis of 3-Ac-NEOS:

[0101] 1) Using T-2 toxin as the reaction substrate, add anhydrous pyridine and acetic anhydride to acetylate the C3 hydroxyl group of T-2 toxin to prepare 3-acetyl-T-2, and pass the reaction product through silica gel column For purification, use n-hexane / ethyl acetate 45:50 as eluent. After the eluate was concentrated, it was identified by mass spectrometry;

[0102] Mass Spectrometry: The m / z of the strongest ion peak detected is 526.66[M+18] + , the molecular weight obtained is 508, which is consistent with the theoretical molecular weight of the target product 3-Ac-T-2 toxin;

[0103] 2) Treat 3-acetyl-T-2 w...

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Abstract

The invention relates to a type A trichothecene toxin polyclonal antibody and its preparation method and application. It uses T-2 toxin as a precursor to carry out structural modification to prepare type A trichothecene toxin The immunoreactive group hapten 3-Ac-NEOS of the mother nucleus structure, and through the coupling with the macromolecular protein BSA and OVA, only the immunoreactive small-molecule hapten is converted into an immunogenic complete antigen, immune For animals, the titer of the prepared antiserum is high, which meets the requirements of immunological detection for antibodies. Based on this antibody, the immunological detection technology established is easy to operate, strong in specificity and high in sensitivity, and can be widely used in food, feed and animal body A-type trichothecenes (including cryptic type A monothecenes) Tectosporine toxins) detection. For example, the ELISA kit prepared with this antibody can be used to detect the hidden T-2 toxin in shrimp.

Description

technical field [0001] The present invention relates to a polyclonal antibody and its preparation method and application. The 3-acetyl group-Neo solani fusarium enol hapten is prepared by structurally modifying the T-2 toxin, and the bridge and carrier protein are constructed by the succinic anhydride method. The combination forms an antigen system suitable for animal immunization, and the polyclonal antibody is obtained after animal immunization, specifically relating to the polyclonal antibody and its preparation method and its application in detecting type A trichothecenes. Background technique [0002] Trichothecenes are a group of mycotoxins with similar chemical properties, which are produced by fungi of the genera Fusarium, Trichoderma, Trichothecene, Cephalosporium, Magnolias, Verticillium and black botrytis, These toxins not only contaminate cereal crops such as wheat, barley, and corn, but also endanger economic crops such as potatoes, and livestock products such a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/14C07K16/06C07K14/765C07K14/77C07D493/10G01N33/543
Inventor 王雅玲吴朝金孙力军邱妹徐德峰刘颖
Owner GUANGDONG OCEAN UNIVERSITY
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