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
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
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 ...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More 