Preparation methods and applications of benzylpenicilloic acid artificial antigen and benzylpenicilloic acid artificial antibody

A technology of penicillin thiazole and penicillin, which is applied in the field of preparation of benzyl penicillin artificial antigen and its antibody, achieving a high degree of similarity and retaining complete effects

Inactive Publication Date: 2014-01-01
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Penicillin G and its degradation product benzyl penic...

Method used

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  • Preparation methods and applications of benzylpenicilloic acid artificial antigen and benzylpenicilloic acid artificial antibody
  • Preparation methods and applications of benzylpenicilloic acid artificial antigen and benzylpenicilloic acid artificial antibody
  • Preparation methods and applications of benzylpenicilloic acid artificial antigen and benzylpenicilloic acid artificial antibody

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preparation example Construction

[0046] The preparation method of benzyl penicillin thiazole acid antibody:

[0047] 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:

[0048] 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;

[0049] Booster immunization: immunize animals with a solution prepared by dissolving 0.5 mg of the above-me...

Embodiment 1

[0054] Preparation method of benzyl penicillin thiazole acid artificial antigen

[0055] With (2S, 5R, 6R)-3,3 dimethyl-6-(2-phenylacetamido)-7-oxo-4-thia-1-azabicyclo[3,2,0]heptane -2-Formic acid, with the amide group CO=NH as a bridge, is connected to hemocyanin (keyhole limpet hemocyanin, KLH) through the active ester method to synthesize artificial antigens; the specific method is:

[0056] 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 4.0 ml 1 M 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.

[0057] Take 0.5000g of the obtained white solid and dissolve it in 2ml of THF, add 0.1500g of N-hydroxysuccinimide and 0.5000g of N,N-dicyclohexylcarbodiimide, and stir the reaction overnight at 22-...

Embodiment 2

[0059] Preparation method of penicillin thiazole acid antibody

[0060] 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:

[0061] 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;

[0062] Booster immunization: immunize animals with a solution prepared by dissolving 0.5 mg of the above-mentioned arti...

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Abstract

The invention discloses preparation methods and applications of a benzylpenicilloic acid artificial antigen and a benzylpenicilloic acid artificial antibody, and relates to a method for simply, quickly and effectively detecting penicillin G and a degradation product of penicillin G, namely benzylpenicilloic acid. The specific antibody to penicillin G and the degradation product of penicillin G, namely benzylpenicilloic acid, is prepared by designing and synthesizing a hapten and the artificial antigen of a penicillin veterinary drug penicillin G with a 6-aminocaproic acid molecular structure and a beta-lactam ring molecular structure, and the degradation product of penicillin G, namely benzylpenicilloic acid; a molecular specific antigenic determinant of the veterinary drug is highlighted; the chemosynthesis difficulty is overcome; and an immune animal induces to generate the specific antibody with very high affinity.

Description

technical field [0001] The invention belongs to the technical field of chemistry, and relates to a preparation method and application of benzyl penicillin thiazole acid artificial antigen and antibody thereof. 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 cows and other edible animals. However, due to irregular drug use and improper collection of milk samples, the phenomenon of penicillin drug residues in food of animal origin is serious. Residues of penicillin drugs in the body will cause severe allergic reactions, and the rate of allergic reactions ranks first among all kinds of drugs. Because penicillin is easily degraded into penicilloic aci...

Claims

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Application Information

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IPC IPC(8): C07K14/795C07K1/107C07K16/44G01N33/543
CPCC07K14/795C07K16/06C07K19/00G01N33/54306
Inventor 张燕王俊平王硕江月明
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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