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Clenbuterol immunogen, coatingen and application thereof in colloidal gold test paper

An immunogen and coating 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 clenbuterol colloidal gold rapid test products currently on the market cannot meet the current situation of national limit requirements

Method used

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  • Clenbuterol immunogen, coatingen and application thereof in colloidal gold test paper
  • Clenbuterol immunogen, coatingen and application thereof in colloidal gold test paper
  • Clenbuterol immunogen, coatingen and application thereof in colloidal gold test paper

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] The synthesis of embodiment 1 clenbuterol hapten

[0040] Weigh 1 mmol of clenbuterol, 5 mmol of 4-bromobutyric acid, and 10 mmol of sodium hydroxide, and reflux in 20 mL of methanol for 48 h. The reaction system was cooled to room temperature, the pH was adjusted to neutral, and the components were evaporated to dryness by rotary evaporation, and the target clenbuterol hapten was separated by using a silica gel column.

[0041] The prepared hapten structure is:

[0042]

Embodiment 2

[0043] Synthesis of Example 2 Clenbuterol Immunogen

[0044] Dissolve 0.1 mmol of the clenbuterol 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 frozen in a -20°C freezer.

[0045] The prepared immune antigen structure is:

[0046]

[0047] (II)

Embodiment 3

[0048] Embodiment 3 experimental animal immunization

[0049] The hapten clenbuterol was used to immunize female balb / c mice for 6 weeks, 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 clenbuterol immunogen, coatingen and application thereof in colloidal gold test paper, and belongs to the technical field of immunology. The immunogen and the coatingen are products of hapten coupling carrier proteins obtained by reaction of clenbuterol and 4-bromobutyric acid. The hapten is similar to the clenbuterol corresponding body on molecular structure, stereo chemistry and electronic distribution. The hapten molecules have active groups convenient for coupling with protein carriers, and the presence of the active groups has no influence on the electronic distribution of molecules of an object to be detected. After the hapten is coupled with hemocyanin, the body can produce clenbuterol-orientated antigens with high titer and good specificity, wherein the antigens are adsorbed on a glass cellulose membrane; and the coatingen constructed after coupling the human albumin can be adsorbed on a nitrocellulose membrane to construct the clenbuterol colloidal gold test paper.

Description

technical field [0001] The invention relates to a clenbuterol immunogen and a coating agent, belonging to the technical field of immunology. Background technique [0002] With global economic integration and internationalization of food trade, food safety has become an important global public health issue. The chemical residues in agricultural products and animal products have obvious acute and chronic toxicity to the human body, including special toxicity such as cancer and chromosomal mutation, which directly endanger the health and life of the people. The extensive use of these drugs also constitutes a direct threat to ecology and livestock and poultry health. Food safety problems not only endanger people's health, affect international trade, cause heavy economic losses, but also lead to social instability. With my country's accession to WTO, our international trade of livestock and poultry products and aquatic products will usher in a more intense competitive environme...

Claims

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Application Information

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Patent Type & Authority Applications(China)
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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