Preparation of egg yolk antibody against citrinin

A technology of citrinicillin-resistant chickens and citrinin-resistant chickens, applied in the field of immunology, can solve the problems of the preparation method and application of egg yolk antibodies that have not been seen in citrinicillin-resistant chickens, and achieve broad application prospects, social and economic benefits, large yields, and specificity high effect

Inactive Publication Date: 2009-05-20
深圳市绿诗源生物技术有限公司
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  • Application Information

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Problems solved by technology

But there is no report on the preparation method and application of anti-ciritillin chicken egg yolk antibody

Method used

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Embodiment 1

[0017] Embodiment one: the preparation of anti-citrinin chicken egg yolk antibody

[0018] (1) Synthesis of citrinin immune antigen:

[0019] Accurately weigh 0.5 mg of citrinin and dissolve in 200 μL of methanol, add 1 mg of carbonyldiimidazole, mix well, react at room temperature for 30 minutes, evaporate to dry naturally, dissolve with 200 μL of DMSO, drop into bovine serum albumin BSA solution, 37 After the reaction, the reaction mixture was placed in a dialysis bag and dialyzed with 0.01mol / L phosphate buffer solution (PBS pH7.4) at 4°C. times, vacuum freeze-dried and stored at -20°C. Dissolve 1 mg of citrinin artificial antigen powder in 1 mL of 0.1 mol / L phosphate buffered saline solution (PBS pH 7.4), add Freund's adjuvant at a ratio of 1:1 (V / V), and fully emulsify to obtain immune antigen.

[0020] (2) Immunization of hens with citrinin immunization antigen:

[0021] Four 22-week-old Laihang hens were taken, and the emulsified citrinin immune antigen was injected in...

Embodiment 2

[0024] Embodiment two: the preparation of anti-citrinin chicken egg yolk antibody

[0025] (1) Synthesis of citrinin-coated antigen:

[0026] Dissolve 0.5 mg of citrinin in 500 μL of dimethylformamide (DMF), add 1 mg of carbodiimide hydrochloride (EDC) and 1 mg of N-hydroxysuccinimide (NHS) successively, mix well, and stir with magnetic force React at room temperature in the dark for 4 hours, then add 2 mL of 5 mg / mL ovalbumin OVA (10 mg OVA dissolved in 2 mL of pH 7.4 0.1 mol / L phosphate buffer) solution drop by drop into the mixture, and avoid light under magnetic stirring React at room temperature for 6 hours. After the reaction, the reaction mixture was placed in a dialysis bag and dialyzed against phosphate buffered saline (PBS) at 4°C. Store at ℃.

[0027] (2) Immunization of hens with citrinin immunization antigen:

[0028] Four 22-week-old Laihang hens were taken, and the emulsified citrinin immune antigen was injected into the subcutaneous, double wings, thigh, and...

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Abstract

The invention discloses a preparation method of an egg yolk antibody against citrinin, comprising the following steps: (1) to prepare the artificial antibody of citrinin; (2) to inject the artificial antibody of citrinin into a hen body to immune; and after 4 times of immunity, to collect eggs continuously for 1 to 2 months; and (3) to extract the egg yolk antibody against citrinin. The prepared egg yolk antibody of anti-citrinin has the advantages of low preparation cost, large productivity, and high specificity. The method can be applied to the development of gold colloid and the establishment of fast examination technology with rapidness, sensitivity, specificity to the citrinin, and has a great application prospect in the safety examination of agricultural products and foods.

Description

technical field [0001] The invention belongs to the technical field of immunology, and in particular relates to a preparation method of an anti-citinillin chicken egg yolk antibody. Background technique [0002] Citrinicillin (CTN) is a mycotoxin produced by certain strains of Penicillium and Aspergillus, which was isolated and purified for the first time in 1931. It can cause significant nephrotoxicity, and has teratogenic and carcinogenic effects. Since citrinin-producing fungi are widely distributed in nature, they often cause mildew in agricultural products, food, and feed. In recent years, the contamination of citrinin in food has attracted more and more attention from all over the world. In 1991, the European Branch of the Natural Toxin Detection Committee of the International Academy of Life Sciences listed it as one of the toxins that must be detected. [0003] Traditional analysis methods of citrinin, such as high performance liquid chromatography, high performan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/44C07K16/02
Inventor 赵肃清林壮森高燕红张焜方岩雄黄宝华
Owner 深圳市绿诗源生物技术有限公司
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