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Artificial antigen and specific antibody of veterinary drug penicillin G degradation product benzylpenicilloic thiazole acid

A technology of penicillin thiazole and artificial antigen, which is applied in the field of design and synthesis of penicillin thiazole small molecular compound artificial hapten and specific antigen, to achieve the effect of complete retention and high similarity

Inactive Publication Date: 2011-05-04
TIANJIN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Penicillin G and its degradation product benzyl penicillin thiazole acid artificial antigen has not been reported

Method used

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  • Artificial antigen and specific antibody of veterinary drug penicillin G degradation product benzylpenicilloic thiazole acid
  • Artificial antigen and specific antibody of veterinary drug penicillin G degradation product benzylpenicilloic thiazole acid
  • Artificial antigen and specific antibody of veterinary drug penicillin G degradation product benzylpenicilloic thiazole acid

Examples

Experimental program
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Effect test

Embodiment 1

[0032] Preparation method of benzicillin thiazole acid artificial antigen

[0033]With (2S, 5R, 6R)-3,3-dimethyl-6-(2-phenylacetamido)-7-oxo-4-thia-1-azabicyclo[3,2,0]heptane Alkane-2-carboxylic acid, with amide group CO=NH as a bridge, is connected to hemocyanin (keyhole limpet hemocyanin, KLH) by active ester method to synthesize artificial antigen; the specific method is:

[0034] Take 1.5 mg of (2S, 5R, 6R)-3,3-dimethyl-6-(2-phenylacetamido)-7-oxo-4-thia-1-azabicyclo[3,2,0 ]heptane-2-carboxylic acid potassium salt was added to 4.0ml 1M hydrochloric acid to obtain the hapten (2S,5R,6R)-3,3-dimethyl-6-(2-phenylacetamido)-7-oxo-4 - Thia-1-azabicyclo[3,2,0]heptane-2-carboxylic acid as a white solid, washed with double distilled water and dried to obtain the hapten.

[0035] Take 0.5000g of the obtained white solid and dissolve it in 2ml of tetrahydrofuran, add 0.1500g of N-hydroxysuccinimide and 0.5000g of N, N-dicyclohexylcarbodiimide, stir and react overnight at 22-25°C, t...

Embodiment 2

[0037] Preparation method of veterinary drug penicillin G degradation product benzyl penicillin thiazole acid antibody

[0038] The immunized animals were female New Zealand white rabbits. The immunization methods were subcutaneous and intramuscular injection. Four booster immunizations were carried out after the initial immunization. The booster immunizations were immunized three times after 2 weeks, 4 weeks and 6 weeks after the initial immunization respectively, and thereafter at intervals of one month. Carry out the fifth immunization, and 9 days later, blood is taken from the ear vein of the rabbit for titer detection. The specific method is as follows:

[0039] Initial immunization: take 1 mg of the artificial antigen synthesized according to the method of claim 3 and dissolve it in a solution prepared by equal volumes of 0.9% NaCl solution and Freund's complete adjuvant, and carry out animal immunization;

[0040] Booster immunization: immunize animals with a solution p...

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Abstract

An artificial antigen and an antibody of a veterinary drug penicillin G degradation product benzylpenicilloic thiazole acid and preparation method thereof. The invention relates to the preparation of a hapten, an artificial antigen and an antibody of benzylpenicilloic thiazole acid having a structure of (2S, 5R, 6R)-3,3-dimethyl-6-(2-pheylacetamino)-7-oxo-4-thia-1-azabicyclo[3,2,0]heptane-2-formic acid and their application in the establishment of immunoassay. The invention solves the problem that traditional physical and chemical analysis methods are complicated, high in cost and slow in analysis, and provides a simple, quick, sensitive and accurate immunoassay technique. According to the invention, (2S, 5R, 6R)-3,3-dimethyl-6-(2-pheylacetamino)-7-oxo-4-thia-1-azabicyclo[3,2,0]heptane-2-formic acid is adopted as a hapten, and the haptens are linked with KLH and HRP respectively to synthesize artificial antigens and enzyme-labeled antigens. The antibody is prepared by animal immunization, blood drawing, antiserum separation, and purification of artificial antigen. The antibody is stable and has good specificity and sensitivity; the synthetic method is simple; the invention is applicable to the rapid immunoassay of veterinary drug penicillin G degradation product benzylpenicilloic thiazole acid, and has a good application prospect.

Description

technical field [0001] The invention belongs to the technical field of immunochemistry and residue analysis of small molecule compounds (molecular weight less than 1000 Daltons); relates to organic synthesis, immunochemistry, biochemistry and physical and chemical testing techniques; in particular, it relates to a compound having a hydrogenated thiazole ring and a β-lactam Design and synthesis of artificial hapten and specific antigen of penicillin veterinary drug penicillin G degradation product benzyl penicillin thiazole acid small molecule compound with ring merged heterocyclic molecular structure. Background technique [0002] Penicillin G (also known as benzyl penicillin) is a kind of penicillin antibiotics. Penicillin antibiotics are the first class of antibiotics to enter clinical application, and they are also the antibiotics with the largest production and usage in my country. Penicillin antibiotics have become the most commonly used drugs to treat diseases of dairy...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/795C07K16/44C07D499/46C07D499/04C07K1/113C12N9/96
Inventor 王硕江月明王俊平张燕
Owner TIANJIN UNIV OF SCI & TECH
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