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Type-A trichothecene polyclonal antibody as well as preparation method and application thereof

A technology of trichothecenes and polyclonal antibodies, which is applied in the direction of anti-fungal/algae/lichen immunoglobulin, animal/human protein, serum albumin, etc., and can solve the problems of complicated operation and limited application range, etc. Achieve high similarity, good specificity and sensitivity, and good immunogenicity

Active Publication Date: 2014-07-30
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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  • Type-A trichothecene polyclonal antibody as well as preparation method and application thereof
  • Type-A trichothecene polyclonal antibody as well as preparation method and application thereof
  • Type-A trichothecene polyclonal antibody as well as preparation method and application thereof

Examples

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 r...

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 ...

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Abstract

The invention relates to a type-A trichothecene polyclonal antibody as well as a preparation method and application thereof. The method comprises the following steps: performing structural modification on T-2 toxin serving as a precursor to prepare a hapten 3-Ac-NEOS with immunoreactivity to an A-type trichothecene mother nucleus structure, coupling the hapten with macromolecular proteins BSA and OVA to convert a micromolecular hapten only having immunoreactivity into a complete antigen with immunogenicity, and immunizing animals. Prepared antiserum is relatively high in titer and the requirement of immunological detection on the antibody is met. An immunological detection technology established on the basis of the antibody is easy and convenient to operate, good in specificity and high in sensitivity, and can be widely applied to detection of type-A trichothecene (including hidden type-A trichothecene) in foods, feeds and animal bodies. For example, an ELISA kit prepared by adopting the antibody can be used for detecting hidden T-2 toxins in prawn bodies.

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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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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