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A kind of anti-zen yolk antibody and preparation method thereof

A yolk antibody, ZEN-BSA technology, applied in the field of immunology, can solve the problems such as anti-ZEN yolk antibodies that have not been seen yet, and achieve the effect of low cost, large output and high specificity

Active Publication Date: 2016-02-10
HENAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in the prior art, there are no related reports about anti-ZEN yolk antibody and its preparation

Method used

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  • A kind of anti-zen yolk antibody and preparation method thereof
  • A kind of anti-zen yolk antibody and preparation method thereof
  • A kind of anti-zen yolk antibody and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] The anti-ZEN egg yolk antibody was prepared by the following method steps:

[0043] 1. Using the Thouvenot method and the carbodiimide method to artificially synthesize ZEN-BSA (zearalenone-bovine serum albumin) antigen; specifically, the following steps are included:

[0044] A. Synthesize ZEN hapten with reference to Thouvenot method

[0045] Dissolve 12 mg of ZEN (Israel fermantek company) in 2 ml of pyridine, add 20 mg of CMO (carboxymethyloxime, carboxymethyl hydroxylamine hemihydrochloride), stir at room temperature for 24 hours, and vacuum dry at 45 ° C. After drying, the residue is dissolved in 6 mL of ultrapure water , use 1mol / L NaOH to adjust the pH to 8.0, then extract with benzene three times, each time 6mL, remove the unreacted ZEN in the water phase, adjust the pH to 3.0 with 1mol / L HCL, and use acetic acid Ethyl ester was extracted 4 times with 10 mL each time. The extract was collected and dried in vacuum to obtain the ZEN-CMO (zearalenone-6-carboxy...

Embodiment 2

[0079] In order to detect the antibody effect of the ZEN yolk antibody prepared in Example 1, the inventors conducted further detection experiments. The experimental process is as follows:

[0080] 1. Determination of the concentration of the sterile anti-ZEN egg yolk antibody extract prepared in step (8) of Example 1

[0081] The concentration of the ZEN egg yolk antibody extract solution after the affinity chromatography column in step (8) of Example 1 was measured, and the absorption value of IgY at 260nm and 280nm after purification by the affinity chromatography column was measured by dual-wavelength ultraviolet spectrophotometry, according to The following formula calculates the IgY concentration:

[0082] IgY concentration (mg / ml) = 1.45 × A 280 -0.74×A 260 .

[0083] The results of ultraviolet spectrophotometry showed that after purification by affinity chromatography, the IgY content was 5.17mg / ml.

[0084] 2. Titer determination of the sterile anti-ZEN egg yolk ...

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Abstract

The invention belongs to the technical field of immunology, and in particular relates to an anti-ZEN (zearalenone) yolk antibody and a preparation method thereof. Anti-ZEN egg yolk antibody is obtained by immunizing laying hens with artificially synthesized ZEN-BSA antigen, collecting eggs, and separating and purifying ZEN egg yolk antibody from egg yolk. The anti-ZEN yolk antibody provided by the present invention is aimed at the ZEN antigen, has high specificity and high titer ratio, and can replace the existing mouse-derived monoclonal antibody; the preparation method of the provided anti-ZEN yolk antibody is compared with the existing The mouse-derived monoclonal antibody has low cost and large yield, which is conducive to the establishment of the ZEN rapid response detection system, so it has positive application significance in the detection of agricultural products and food safety.

Description

technical field [0001] The invention belongs to the technical field of immunology, and in particular relates to an anti-ZEN (zearalenone) yolk antibody and a preparation method thereof. Background technique [0002] ZEN (zearalenone, zearalenone), also known as F-2 toxin, is a 2,4-dihydroxybenzoic acid lactone compound with high estrogen activity. It is mainly caused by infecting corn, wheat, barley, A non-steroidal estrogenic mycotoxin produced by Fusarium graminearum and Fusarium yellow in crops such as oats and sorghum, which can produce various toxic and side effects on humans and animals, for example, it has strong reproductive toxicity and teratogenic effects , can cause hyperestrogenism in animals, lead to infertility or abortion, have a greater impact on pigs, poultry and ruminants, and bring great economic losses to animal husbandry; in addition, ZEN also has cytotoxicity, immunotoxicity, liver Toxicity and potential carcinogenicity, etc., seriously threaten human ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/14C07K16/02
Inventor 刘英杰陈娟黄永伟刘云
Owner HENAN UNIVERSITY