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

Active Publication Date: 2016-09-28
CHENGDU ANTIK BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, in order to ensure the immune protection effect, quadrivalent influenza has been mentioned many times in WHO reports in recent years. The existing trivalent influenza virus chicken egg yolk antibody cannot block the virus of another strain of B (B) influenza virus. and preventive protection
[0007] At the same time, when preparing yolk antibodies, although the immunized chickens are healthy laying hens that have been selected, inspected and vaccinated against certain diseases, there are still some chicken-derived bacteria or / and viruses mixed in the eggs to enter Possibility in yolk antibody, such as Escherichia coli, Salmonella, chicken egg drop syndrome virus, chicken avian leukemia virus, etc., thus affecting the safety of the antibody
At present, some studies use toxic formaldehyde, divinylamine (BEI) and β-propiolactone (BPL) as inactivators, which have inactivation effects on exogenous viruses in egg yolk antibodies, but have a negative effect on the activity of egg yolk antibodies. affect the effect of the effect
The exogenous virus inactivation methods currently used add toxic substances, which have a significant impact on the biological activity of egg yolk antibodies and have potential toxic side effects on the human body

Method used

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  • Chicken egg-yolk antibody capable of resisting influenza virus and preparation method thereof
  • Chicken egg-yolk antibody capable of resisting influenza virus and preparation method thereof

Examples

Experimental program
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Effect test

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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Abstract

The invention discloses a chicken egg-yolk antibody capable of resisting influenza virus. The chicken egg-yolk antibody is used for specifically resisting tetravalence influenza virus. The influenza virus includes influenza A virus subtype H1N1, influenza A virus subtype H3N2, influenza B virus subtype Victoria series and influenza A virus subtype Yamagata series. The preparation method of the chicken egg-yolk antibody comprises the following steps of 1, preparing a tetravalence influenza virus antigen; 2, performing immunity on hens; 3, collecting immunized eggs laid by the immunized hens; 4, preparing a mixed antibody stock solution from the immunized eggs; 5, detecting titer of the antibody stock solution, and a stabilizer is added cooperatively, so that the chicken egg-yolk antibody capable of resisting tetravalence influenza virus is obtained. The chicken egg-yolk antibody can be combined with the tetravalence influenza virus specificity pathogen for a short period of time, the tetravalence influenza virus can be effectively killed for a long period of time, the safety performance is good, no toxic or side effect is generated, and the chicken egg-yolk antibody can be effectively used for preventing pathogen corresponding to human pharyngeal and nasopharynx mucosas.

Description

technical field [0001] The invention relates to the technical field of biological antibody immunization, in particular to an anti-influenza virus chicken egg yolk antibody and a preparation method thereof. Background technique [0002] Influenza virus, referred to as influenza virus, is an RNA virus that causes influenza in humans and animals. In taxonomy, influenza virus belongs to Orthomyxoviridae. Under the electron microscope, influenza virus is spherical or oval, with a diameter of about 80~120nm, filaments can be seen in the first separation, with different lengths. According to the antigenicity of viral nucleoprotein (NP) and matrix protein (MA), it can be divided into A, B and C. Influenza A has different subtypes according to the antigenicity of surface glycoproteins HA and NA. Currently, there are 16 HA subtypes (H1-H16) and 9 NA subtypes (N1-N9). The viral genome consists of 8 segmented single-stranded negative-sense RNAs. Among them, 8 gene fragments of influe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/10C07K16/02C07K1/36C07K1/34C07K1/30
CPCC07K16/02C07K16/1018C07K2317/11C07K2317/23C07K2317/31
Inventor 林小军朱玲林烨暄万强雷雨冯梅胡方焱何珊江秀坤叶琳
Owner CHENGDU ANTIK BIOTECH CO LTD
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