Chicken egg-yolk antibody capable of resisting influenza virus and preparation method thereof
A technology of chicken yolk antibody and anti-influenza virus, applied in the field of anti-influenza virus chicken yolk antibody and its preparation, can solve the problem that the virus cannot be blocked and preventive protection, the biological activity of egg yolk antibody is obviously affected, and the activity of egg yolk antibody is affected, etc. problem, to achieve the effect of less allergic reaction, good curative effect and stable biological activity
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preparation example Construction
[0047] Its specific preparation method comprises the following steps:
[0048] (1) Take type A, type B and type C influenza virus strains and inoculate chicken embryos respectively; culture the chicken embryos inoculated with different types of influenza virus strains; when the chicken embryos have lesions, collect the virus liquid respectively , followed by virus inactivation; finally, four-valent antigens were prepared after ultrafiltration concentration and sucrose density gradient centrifugation;
[0049] (2) Mix the prepared tetravalent antigen 1:1:1:1, add an equal volume of Freund's complete adjuvant or Freund's incomplete adjuvant and mix, then resonate on a tissue lysis instrument at 25Hz for 5min, repeat twice, The emulsified mixed solution is obtained, and after passing the safety test, sterility test and physical property inspection, it is used for immunized laying hens; take a healthy laying hen of appropriate age, and take the emulsified mixed solution in the del...
Embodiment 1
[0067] In order to verify the performance of egg yolk antibody inactivation virus in this embodiment, the following tests were carried out:
[0068] Take the diluted influenza A H1N1, A H3N2, B (Victoria series), and B (Yamagata series) influenza virus suspensions and mix them with the same amount of samples to be tested. In a 96-well cell plate grown into a single layer (each well contains 100ul of mixed solution and 100ul of virus culture solution), placed in a CO2 incubator at 33°C for 48 hours, then take 50ul of cell suspension from each well and mix with 50ul of 1% guinea pig red blood cell suspension , placed at room temperature for 45 minutes to observe the results. The average inactivation logarithm value (TCID50) of the product to be tested against the four types of influenza virus was calculated by the endpoint dilution method.
[0069] At room temperature, after 3 repeated experiments, the average inactivation logarithms of the samples to be tested against influenz...
Embodiment 2
[0072] In this example, on the basis of the above examples, in order to verify that the anti-influenza virus chicken egg yolk antibody has a long residence time in different parts of living organisms, the obtained anti-influenza virus chicken egg yolk antibody was subjected to drug residence time small animal imaging Experiment, the specific process is as follows:
[0073] Adjust the pH of the buffer system (sodium bicarbonate buffer system) to 8.5-10.0, add a small amount of fluorescent dye to the anti-influenza virus chicken egg yolk antibody solution, and mix vertically overnight at 4°C. Then ultrafiltration centrifugation (5000-6000g, centrifugation time 4-6min) to remove unreacted fluorescent molecules.
[0074] Test method: The anti-influenza virus chicken egg yolk antibody stock solution containing fluorescent molecules is made into oral spray. Divided into three experimental groups: group a: fasting for 8 hours after spraying, no fasting; group b: fasting for 8 hours ...
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