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Valnemulin artificial antigen and preparation method and application thereof

A technology of venomulin and reaction, which is applied in the field of venomulin artificial antigen and its preparation, can solve the problems of unproven results and short clinical application time, and achieve broad application prospects, simple sample pretreatment, and high accuracy high effect

Active Publication Date: 2012-10-24
北京明日达科技发展有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although studies have reported that warnemulin is less prone to drug resistance compared with other drugs, its clinical application time is not long, and it cannot prove its results after long-term use

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1. Preparation and identification of vornemulin artificial antigen

[0033] Vanimulin: purchased from Sigma Aldrich; product catalog number: 32971.

[0034] 1. Preparation of Vonemulin Artificial Antigen

[0035] Dissolve 50mg Valnemulin (Valnemulin) and 100mg carrier protein (bovine serum albumin BSA) in 2ml 0.01M pH7.4 PBS to obtain solution I; weigh 100mg carbodiimide (EDC) and dissolve it in 1ml In water, solution II is obtained; solution II is added to solution I and reacted for 2 hours at 20° C. to obtain the reaction product (Vonemulin artificial antigen). The reaction product was dialyzed, and the conditions of the dialysis were: 4° C., dialysis with 0.01M pH 7.4 PBS for 72 hours. After dialysis, the compound was centrifuged at 5000-8000 rpm at 4°C for 30 minutes, and the supernatant (Vonemulin artificial antigen) was taken, aliquoted into ampere bottles, and stored at -20°C.

[0036] 2. Identification of Vonimulin Artificial Antigen

[0037] The supernatant (V...

Embodiment 2

[0038] Example 2. Preparation and specific identification of antiserum

[0039] Vonimulin standard: purchased from Sigma Aldrich; product catalog number: 32971.

[0040] New Zealand white rabbit: purchased from Beijing Experimental Animal Research Center.

[0041] HRP-Goat anti-rabbit IgG: purchased from Shanghai Ruicong Technology Development Co., Ltd.; product catalog number: KT117.

[0042] 1. Preparation of Vonimulin Antiserum

[0043] 100 New Zealand white rabbits were randomly divided into an experimental group and a control group (50 in each group); the experimental group was immunized with the Vonemulin artificial antigen prepared in step 2, and the control group was immunized with bovine serum albumin (BSA); each Rabbits are immunized at 1 mg / mL (based on the amount of BSA) every two weeks for a total of 7 immunizations; 7 days after the third immunization, 7 days after the fourth immunization, and 7 days after the fifth immunization , 7 days after the sixth immunization and ...

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Abstract

The invention discloses a valnemulin artificial antigen and a preparation method and application thereof. The valnemulin artificial antigen is a compound as shown in a formula I, wherein n denotes a natural number from 1 to 25; and K denotes carrier protein. When the artificial antigen is used to immunize a New Zealand white rabbit, an antibody with specific reaction to valnemulin can be obtained. According to ELISA experiments, the titer of the obtained antiserum reaches 1:102400. The invention provides a wide application and development space for the development of detection reagent used inthe enzyme-linked immunoassay method of the valnemulin and the preparation of the enzyme-linked immunoassay reagent kit of the valnemulin. When the antibody obtained by using the artificial antigen is used for the detection of the valnemulin, the operation is simple and convenient, the specificity is good and the accuracy is high. The antibody can be used for the rapid and high-efficiency screening of the valnemulin in animal-based foods and a technical foundation is laid for the rapid detection of the valnemulin. The formula I is as shown is the accompanying drawing.

Description

Technical field [0001] The invention relates to an artificial antigen of Vonimulin and its preparation method and application. Background technique [0002] Valnemulin (Valnemulin) is a new generation of pleuromu.tilin (pleuromu.tilin) ​​semi-synthetic antibiotics. It belongs to diterpenes and is a similar drug to tiamulin. The raw material of Vonemulin that is now on the market is Vonemulin hydrochloride, with the chemical name {[2~((R)-2-((R)-2—amino—3-methylbutyrylamino))—1,1—dimethylethyl Group] mercapto}acetic acid (3as, 4R, 5S, 6S, 8R, 9R, 9aR, IOR)-octahydro-5, 8-dihydroxy-4, 6, 9, 10-tetramethyl-6-vinyl- 3a,9-propane-3aH-cyclopentacyclooctene-1(4H)-ketone-8-ester hydrochloride, molecular formula C3lH53CINz 05S, relative molecular weight 601.30, chemical structure formula see formula (II). Vonimulin hydrochloride is a white or light yellow non-crystalline powder with hygroscopicity; it is easily soluble in water and absolute ethanol, and almost insoluble in tert-butyl me...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/765C07K14/77C07K14/795C07K1/10C07K16/44G01N33/577G01N33/53
Inventor 江海洋王战辉吴小平徐飞史为民米铁军李娜王亚辉张静
Owner 北京明日达科技发展有限责任公司
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