Albendazole artificial antigen and preparation method and application thereof

A technology of albendazole and antigen, which is applied in the field of albendazole artificial antigen and its preparation, can solve the problems of cumbersome operation steps, inapplicability to large-scale sample screening and detection, and high cost, and achieve high purity, high yield, The effect of simple synthesis method

Active Publication Date: 2016-10-12
北京维德维康生物技术有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these methods have strong specificity and high sensitivity, the sample pretreatment steps are cumbersome and costly, and they are not suitable for the screening and detection of large batches of samples.

Method used

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  • Albendazole artificial antigen and preparation method and application thereof
  • Albendazole artificial antigen and preparation method and application thereof
  • Albendazole artificial antigen and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Embodiment 1, the preparation of albendazole hapten

[0041] One, the preparation of albendazole hapten

[0042] Add 500mg of albendazole and 20ml of dimethylformamide (DMF) (as the solvent for the reaction) into a 50ml round-bottomed flask, dissolve it, add 580mg of 4-aminobutyric acid, react at 80°C for 5h, TLC detects that the raw materials are processed after the reaction is completed , the reaction solution was added dropwise to 10ml of ice water, a white solid was precipitated, filtered, washed with water and dried to obtain 320mg of the product.

[0043] The reaction equation is as follows:

[0044]

[0045] 2. Structural identification of albendazole hapten

[0046] Mass spectrometry detection is carried out to gained 320mg product, M+:M+H=337.2, M+Na=359.2, M-:M-H=335.4, as figure 1 shown. The results show that its chemical structure is as shown in Formula I (MW=336.4), which is the albendazole hapten.

[0047]

Embodiment 2

[0048] Embodiment 2, preparation and structural identification of albendazole artificial antigen

[0049] 1. Preparation of albendazole artificial antigen

[0050] 1. Synthesis of immunogen

[0051] (1) Dissolve 15 mg of the hapten (Formula I) prepared in Example 1 in 1.5 mL of dimethylformamide (DMF). After dissolving completely, add 1-(3-dimethylaminopropyl)-3 - Ethylcarbodiimide hydrochloride (EDC) 26mg, N-hydroxysuccinimide (NHS) 25mg, room temperature (20-25°C) magnetic stirring reaction for 3h, to obtain solution I.

[0052](2) Weigh 50 mg of bovine serum albumin (BSA), dissolve it in 3.5 mL of 0.1 M carbonic acid buffer, stir at 400 rpm for 10 min, and fully dissolve to obtain solution II.

[0053] Wherein, the pH of the 0.1M carbonic acid buffer is 9.6, the solvent is water, and the solute and its concentration are as follows: Na 2 CO 3 1.59g / L, NaHCO 3 2.94g / L.

[0054] (3) Take the above solution I, add it dropwise to the above solution II in an ice-water bath...

Embodiment 3

[0064] Embodiment 3, albendazole artificial antigen immunization animal prepares antiserum

[0065] 1. Animal immunity

[0066] New Zealand white rabbits were immunized with the albendazole artificial antigen "albendazole-BSA" obtained in step Example 2 as an immunogen. The dose of each immunization is 100-200 μg, and the immunization method is multi-point subcutaneous injection on both shoulders and hind thighs, and about 1 / 4 of the immunogen is used in each area. For the first immunization, dilute the immunogen with normal saline, then mix it with Freund's incomplete adjuvant 1:1 (volume ratio) to make an emulsifier, and take the same dose of immunogen plus an equal volume of Freund's incomplete adjuvant every 2 weeks After mixing and emulsifying the vaccine, the immunization was boosted once. After 3 times of immunization in this way, the same dose of immunogen was added with Freund's incomplete adjuvant for the last immunization at an interval of 3 to 4 weeks. Blood was c...

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Abstract

The invention discloses an albendazole artificial antigen and a preparation method and an application thereof. The albendazole artificial antigen provided by the invention is an antigen obtained by coupling albendazole hapten represented by the formula I with a vector protein. The albendazole artificial antigen provided by the invention has the advantages of simple synthetic method, high purity and high yield, and has great value to preparation of the albendazole antibody and detection of albendazole drug residues.

Description

technical field [0001] The invention belongs to the fields of biomedicine and chemistry, and relates to an albendazole artificial antigen and a preparation method and application thereof. Background technique [0002] Albendazole is a broad-spectrum anthelmintic with high efficiency and low toxicity. Clinically, it can be used to drive away roundworms, pinworms, tapeworms, whipworms, hookworms, and strongyloid nematodes. After being metabolized into sulfoxides or sulfones in the body, it can inhibit the absorption of glucose by parasites, resulting in the depletion of glycogen in the parasites, or inhibit the fumarate reductase system, hinder the production of ATP, and make the parasites unable to survive and reproduce. Residues in animal food are a potential threat to human health. [0003] At present, the commonly used methods for the detection of veterinary drug residues include gas chromatography, high performance liquid chromatography, and gas chromatography-mass spec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D235/30C07K14/765C07K14/77C07K16/44G01N33/543
CPCC07D235/30C07K14/765C07K14/77C07K16/44C07K19/00G01N33/543
Inventor 刘志萍苏丽芳于书英吴小平王文珺温凯秦誉王照鹏杨柳邢维维
Owner 北京维德维康生物技术有限公司
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