Preparation method of anti-influenza A H1N1 virus specific IgY and related formulation thereof

An influenza virus, avian influenza virus technology, applied in the preparation method of peptide, antiviral agent, antiviral immunoglobulin, etc., can solve the problem of unstoppable H1N1 influenza virus, central nervous system side effects, failure to detect, etc. problems, to achieve the effect of good immune conditioning and inhibition, and elimination of viral infections

Inactive Publication Date: 2009-10-14
深圳雅臣生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] 3. Wear a mask: Because the H1N1 influenza virus is as small as the general influenza virus, the general mask cannot block the H1N1 influenza virus at all.
On April 30, 2009, the flight from the epidemic area Mexico to Shanghai had a H1N1 influenza virus carrier on board. After two hours of body temperature testing at Shanghai Airport; however, such strict multiple body temperature testing; still failed any abnormality detecte

Method used

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  • Preparation method of anti-influenza A H1N1 virus specific IgY and related formulation thereof
  • Preparation method of anti-influenza A H1N1 virus specific IgY and related formulation thereof
  • Preparation method of anti-influenza A H1N1 virus specific IgY and related formulation thereof

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Experimental program
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Example Embodiment

[0061] 7. Preparation of crude extract

[0062] First, according to the different birds to be immunized and the antigen used for immunization, the immunized eggs are classified and coded. Wash the immune eggs with running water, scrub and disinfect with alcohol, break the immune eggs with a whisk, filter out the egg whites with a yolk sieve, leave the egg yolk, and stir well; add distilled water 4-6 times the volume of the egg yolk, dilute and mix Evenly, adjust the pH to 5.5-6.0 with 1.0N HCI solution.

[0063] Stir the diluted solution with adjusted pH value thoroughly, then cool it to 2-6°C and let it stand for 12-24 hours; centrifuge the diluted solution at 10,000rpm for 20 minutes; take the supernatant obtained from the separation and ultrafiltration Concentrate 10-20 times by ultrafiltration in the vessel; then add sodium alginate solution and stir until turbidity appears; then add 1.0% CaCl 2 Stir evenly, let stand at 4°C for 8-12 hours; centrifuge at 8,000rpm for 20 minute...

Example Embodiment

[0083] Experimental example 1 :Detection of specific IgY against variant A H1N1 influenza virus and specific IgY against influenza secondary infection bacteria

[0084]Using SDS-PAGE (sodium dodecyl sulfate-polypropylene gel) electrophoresis method, the different anti-mutant H1N1 influenza virus specific IgY and anti-influenza secondary infection bacteria prepared according to the above process were specific The crude IgY extract was tested, and the result was 45-52% IgY. After the crude IgY extract is passed through the column twice, pure IgY is obtained. After SDS-PAGE analysis, the purity reached PAGE purity, as shown in the following table:

[0085] IgY pure product

Example Embodiment

[0086] Experimental example 2 :Detection of the activity of specific IgY against variant A H1N1 influenza virus and bacterial specific IgY against influenza secondary infection

[0087] The anti-mutant H1N1 influenza virus specific IgY and the anti-influenza secondary infection bacteria specific IgY prepared by using the process technology invented by the present inventors are used for activity detection using the "ELISA method" (enzyme-linked immunosorbent assay).

[0088] The test results showed that the prepared two different anti-mutant H1N1 influenza virus specific IgY, of which the binding titer of type I to influenza A H1N1 virus reached more than 1:1024. The antibody binding titer of type II to influenza A H1N1 virus also reached more than 1:512. Similarly, the anti-influenza secondary infection bacteria specific IgY against representative pneumococcus, A cluster B hemolytic streptococcus, Staphylococcus aureus, Haemophilus influenzae antibody titers are also more than 1...

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Abstract

The invention discloses a preparation method of an anti-influenza A H1N1 virus specific IgY and related formulations thereof. The method comprises the steps of preparing a plurality of types of complex antigens aiming at influenza A H1N1, using the complex antigens for immunizing laying hen, utilizing the immune egg yolk for preparing specific IgY crude extract dry powder, conducting purification, removing all types of bacterial virus by filtration, and then preparing the anti-variant influenza A H1N1 virus specific IgY, anti-influenza secondary infection bacterial specific IgY, or anti-variant influenza A H1N1 virus specific complex IgY. In addition, the invention makes the anti-variant influenza A H1N1 virus specific IgY into nanoliposomes, and the obtained nanoliposome anti-variant influenza A H1N1 virus specific IgY enhances healing efficacy. The anti-influenza A H1N1 virus specific IgY can be utilized for preparing various formulations which then can prevent and teat the influenza A H1N1 of human and animals fundamentally; and the invention is more convenient, more economic, safer and more effective than the control method used at the present time.

Description

technical field [0001] The invention relates to an antibody and a novel preparation thereof, in particular to a method for preparing anti-H1N1 influenza virus-specific IgY and related preparations. Background technique [0002] The influenza that first broke out in Mexico in April 2009 and then spread rapidly to many countries and regions in the world was caused by a type A H1N1 [A(H1N1)] influenza virus. This type A H1N1 influenza virus is a mixture of human influenza virus, North American avian influenza virus, and North American, European and Asian swine influenza virus, including human influenza virus, North American avian influenza virus, and North American, European and Asian swine influenza virus. The genetic component of the Asian swine flu virus is a new type of virus with genetic mutations. Most worryingly, this novel virus has spread from person to person. According to the similar characteristics of blood relatives between pigs and humans, it shows that the immu...

Claims

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Application Information

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IPC IPC(8): C07K16/10C07K16/02C07K1/14C07K16/12A61K39/42A61K39/40A61K9/127A61P31/16
Inventor 包晟杨荣鉴包海威
Owner 深圳雅臣生物科技有限公司
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