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Immunogen and coatinggen of chloramphenicol and application thereof in collaurum test paper

A chloramphenicol, immunogen technology, applied in the field of immunology, can solve the problem that the detection limit cannot meet the national limit requirements, and achieve the effect of improving immunogenicity

Active Publication Date: 2011-03-30
深圳市三方圆生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the detection limit of chloramphenicol colloidal gold rapid test products currently on the market cannot meet the current situation of national limit requirements

Method used

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  • Immunogen and coatinggen of chloramphenicol and application thereof in collaurum test paper
  • Immunogen and coatinggen of chloramphenicol and application thereof in collaurum test paper
  • Immunogen and coatinggen of chloramphenicol and application thereof in collaurum test paper

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Experimental program
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Effect test

Embodiment 1

[0038] The synthesis of embodiment 1 chloramphenicol hapten

[0039] Weigh 1 mmol of chloramphenicol, 10 mmol of potassium permanganate and reflux reaction in 20 mL of distilled water for 48 h. Cool the reaction system to room temperature, evaporate excess water by rotary evaporation, add 10 mL of ethyl acetate, and filter to obtain the filtrate. Add 20mL of saturated sodium bicarbonate solution and shake fully. The ethyl acetate layer was discarded. The aqueous phase was adjusted to be neutral, extracted three times with 60 mL of ethyl acetate, and the organic solvent was evaporated to obtain a yellow powdery solid.

[0040] 0.1 g of the yellow solid prepared above was used, dissolved in 2 mL of DMF, and 60 mg of DCC and 40 mg of NHS were added with stirring. After 24 hours of reaction, 0.2 g of 4-aminobutyric acid was added, and the reaction was continued for 48 hours. Filter and retain the filtrate, and use a silica gel column to separate the target chloramphenicol hapt...

Embodiment 2

[0043] Synthesis of embodiment 2 chloramphenicol immunogen

[0044] Dissolve 0.1 mmol of chloramphenicol hapten in 2 mL of DMF, and add 27.5 mg of DCC and 14.4 mg of NHS with stirring. Magnetic stirring was carried out overnight at 4°C. After centrifugation, the supernatant was liquid A, and 140 mg of hemocyanin (KLH) was weighed and dissolved in 10 mL of PBS (pH 8.0) with a concentration of 0.1 mol / L. Add 1 mL of DMF, stir and dissolve to prepare liquid B. Under magnetic stirring, liquid A is gradually dropped into liquid B, and react at 4°C for 12 hours. After centrifugation, the supernatant was taken, dialyzed with normal saline at 4°C for 3 days, and the dialysate was changed 3 times a day. The obtained whole antigen was dispensed into 0.5 mL centrifuge tubes at a concentration of 1 mg / mL. Store in -20°C freezer.

[0045] The prepared immune antigen structure is:

[0046]

Embodiment 3

[0047] Embodiment 3 experimental animal immunization

[0048] The hapten chloramphenicol immunization antigen was used to immunize female balb / c mice for 6 weeks respectively, 3 mice in each group. For the first immunization injection, 100 μL of 100 μg / mL immune antigen was fully emulsified with the same amount of complete Freund’s adjuvant, and injected directly into the intraperitoneal cavity. After an interval of two weeks, take the sampled antigen, emulsify it with 100 μL of incomplete adjuvant, and inject it in the same way.

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Abstract

The invention relates to immunogen and coatinggen of chloramphenicol and application thereof in collaurum test paper, and belongs to the technical field of immunology. The immunogen and the coatinggen are a product of hapten coupling carrier protein obtained by oxidizing the chloramphenicol through potassium permanganate. The hapten is similar to a chloramphenicol counterpart in the aspects of molecular structures, stereochemistry and electron distribution; and hapten molecules have active groups which are convenient to couple with a protein carrier, and the active groups do not influence the electron distribution of analyte molecules. After hemocyanin is coupled by the hapten, organisms can generate antibodies with high valence and good specificity aiming at the chloramphenicol, and the antibodies are adsorbed to glass cellulose membranes; and the coatinggen constructed by coupling human serum protein can be adsorbed to nitrocellulose membranes and is used for constructing the chloramphenicol collaurum test paper.

Description

technical field [0001] The invention relates to a chloramphenicol immunogen and a coater, belonging to the technical field of immunology. Background technique [0002] Chloramphenicol was originally extracted from the culture solution of Streptomyces venezuela, and is now mass-produced by chemical synthesis. Is a class of broad-spectrum antibiotics. It has inhibitory effect on Gram-positive and Gram-negative bacteria. Among them, the effect on typhoid bacillus, influenza bacillus and pertussis bacilli is stronger than other antibiotics, and it is also effective against rickettsial infections such as typhus, but its effect on gram-positive cocci is not as good as that of penicillin and tetracycline. [0003] In the first half of 2002, a series of trade disputes arose between China and the European Union over animal-derived foods. On January 28, 2002, the European Union announced a complete ban on the import of animal-derived food from China on the grounds of chloramphenico...

Claims

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Application Information

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IPC IPC(8): C07K14/795C07K14/765G01N33/558
Inventor 钟松清谭攀李细清张世伟
Owner 深圳市三方圆生物科技股份有限公司
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