Toxoflavin hapten, artificial antigen, antibody, synthesis method and application thereof

A technology of artificial antigen and synthesis method, applied in the field of immunochemical analysis, to achieve the effects of simple reaction operation, easy availability of raw materials, high purity and yield

Inactive Publication Date: 2019-10-01
SHENZHEN CENT FOR DISEASE CONTROL & PREVENTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, there is no relevant report on toxin hapten in China

Method used

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  • Toxoflavin hapten, artificial antigen, antibody, synthesis method and application thereof
  • Toxoflavin hapten, artificial antigen, antibody, synthesis method and application thereof
  • Toxoflavin hapten, artificial antigen, antibody, synthesis method and application thereof

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Experimental program
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preparation example Construction

[0037] Specifically, the synthetic method of the toxin hapten of the above structure comprises the following steps:

[0038] S1. methylhydrazine reacts with compound 1 to obtain compound 2, which has the structural formula The compound 2 has the structural formula

[0039] In this step, methylhydrazine and compound 1 undergo a substitution reaction, the principle of which is known to those skilled in the art through technical manuals or through conventional experimental methods. The present invention does not specifically limit the specific operation of the reaction. Those skilled in the art Regular adjustments can be made according to actual needs.

[0040] S2. Under acidic conditions, the compound 2 reacts with formaldehyde to obtain compound 3, which has the structural formula

[0041] In this step, compound 2 and formaldehyde undergo a carbonylamide condensation reaction under acidic conditions to form a carbon-nitrogen double bond. The reaction principle of the ca...

Embodiment 1

[0061] Embodiment 1: Synthesis of toxin hapten

[0062] S1. methylhydrazine reacts with compound 1 to obtain compound 2, which has the structural formula The compound 2 has the structural formula

[0063] S2. Under acidic conditions, the compound 2 reacts with formaldehyde to obtain compound 3, which has the structural formula

[0064] S3. In the presence of sodium nitrite and acetic acid, the compound 3 is reacted to obtain compound 4, which has the structural formula

[0065] S4. The compound 4 is hydrolyzed to obtain the toxin hapten.

[0066] Wherein, the synthetic method of the compound 1 is: (1) provide the compound A, the molecular formula of the compound A is NH 2 (CH 2 ) n COOEt; (2) the compound A reacts with trimethylsilyl isocyanate to obtain compound B, which has the structural formula The compound B reacts with malonic acid and acetic anhydride to obtain compound C, which has the structural formula The compound C is reacted with phosphorus oxychlo...

Embodiment 2

[0067] Embodiment 2: the synthesis of toxin immunization antigen

[0068] Get 20 mg of carrier protein (bovine serum albumin, BSA) and dissolve it in 4 ml of phosphate buffer (pH7.4), add 5 mg of toxin hapten, mix well, add 10 mg of EDC (1-(3-dimethylaminopropyl base)-3-ethylcarbodiimide hydrochloride), react at room temperature (25°C) for 2 hours, dialyze to remove small molecules that are not bound to the carrier protein, and collect the dialyzed protein as the toxin immune antigen.

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PUM

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Abstract

The invention discloses a preparation method of a toxoflavin hapten and a toxoflavin artificial antigen, and the toxoflavin artificial antigen can be used for immunizing animals to obtain antibodies with high specificity and high sensitivity. The antigen and antibodies prepared by the method and sample treatment scheme can provide a simple and quick technical approach for highly sensitive detection of toxiflavin.

Description

technical field [0001] The invention relates to the technical field of immunochemical analysis, in particular to toxin haptens, artificial antigens, antibodies and synthesis methods and applications thereof. Background technique [0002] Yeasted rice noodle poisoning is a type of bacterial food poisoning with a very high fatality rate in the vast rural areas of our country. Pseudomonas cocos subspecies fermented rice noodle is the cause of fermented rice noodle food poisoning and deteriorating white fungus poisoning. Studies have proved that the pathogenic substance of this fungus is two toxins produced during its growth and reproduction: bongkrekic acid (bongkrekic acid) acid, BA) and toxoflavin (toxoflavin, TF). The two toxins are small molecular substances of fatty acids, which are highly toxic, heat-resistant, and highly toxic, and cannot be destroyed by general cooking methods. Therefore, contaminated food can easily cause poisoning. The production of zymosic acid is ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D487/04C07K14/765C07K14/77C07K14/795C07K16/44G01N33/53G01N33/532G01N33/558G01N33/58
CPCC07D487/04C07K14/765C07K14/77C07K14/795C07K16/44C07K19/00G01N33/53G01N33/532G01N33/558G01N33/587
Inventor 柯跃斌李金峰黎诚耀丁晔赵芳肖云军刘博超
Owner SHENZHEN CENT FOR DISEASE CONTROL & PREVENTION
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