Antigen-antibody complex for preventing and/or treating bird flu

A technology of antigen-antibody and complex, applied in the direction of antibodies, antibody medical components, antiviral agents, etc., can solve the problems of gene antigen drift, antigen variation, etc., and achieve strong immune response, strong response, simple and fast production

Active Publication Date: 2011-12-28
INST OF ZOOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Avian influenza viruses are composed of 10 proteins encoded by 8 segments of RNA, among which hemagglutinin (HA) and neuraminidase (NA) are the main surface antigens, but their genes are prone to antigenic drift and antigenic variation, so the above-mentioned several Neither method is applicable

Method used

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  • Antigen-antibody complex for preventing and/or treating bird flu
  • Antigen-antibody complex for preventing and/or treating bird flu

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Preparation of avian influenza antigen-antibody complex preparation

[0026] The specific preparation method is operated as follows:

[0027] (1) Inoculate 9-day-old SPF chicken embryos with avian influenza H5N1 subtype virus strain, collect viral allantoic fluid under aseptic conditions after propagation, add 0.4% formaldehyde, and inactivate at 37°C for 72 hours.

[0028] (2) Purified avian influenza virus from blood source (healthy chicken red blood cells treated with normal saline containing formaldehyde). The concentration of the avian influenza virus was determined to be 1.38 mg / ml after 20-fold dilution of the antigen and 1.22 mg / ml after the antibody was diluted 20-fold. The antibody is IgY prepared after multiple immunizations with a conventional recombinant avian influenza virus H5 subtype bivalent inactivated vaccine (H5N1, Re-1 strain + Re-4 strain);

[0029] (3) Dilute the antigen of the above concentration by 100 times and the antibody by 150 times, and...

Embodiment 2

[0031] Example 2: Immune response and curative effect caused by avian influenza antigen-antibody complex preparation

[0032] (1) Materials and methods

[0033] A. Antigen: The whole virus of avian influenza H5N1 virus strain which is inactivated by formaldehyde and purified by formaldehyde saline is the antigen.

[0034] B. Antibody: The multivalent antibody IgY prepared by immunizing healthy laying hens with eggs produced by inactivated vaccines of H5 subtype of avian influenza as the antibody.

[0035] C. In the ELISA square titration test, the purified influenza virus is used as the antigen, IgY is the antibody, and biphenyldiammonium (OPD) is the substrate. Reading at 490nm, the absorbance value is 0.4 as the best ratio, and the antigen is diluted The concentration after 20 times is 1.38 mg / ml, and the concentration of antibody after 20 times dilution is 1.22 mg / ml, which is used for the preparation of antigen-antibody complexes. Dilute the antigen of the above concentration by ...

Embodiment 3

[0038] Example 3: Immune response and curative effect caused by avian influenza subunit complex preparation

[0039] (1) Materials and methods

[0040] A. Antigen: RNA is extracted from chicken embryo allantoic fluid of avian influenza virus, RT-PCR is performed, HA target gene is recovered, connected to pET-21b vector, and cloned and expressed in BL21 competent cells. The bacteria were lysed by ultrasonic, and HA protein was detected by SDS-PAGE electrophoresis. Purify protein on nickel column.

[0041] B. Antibody: The multivalent antibody IgY prepared by immunizing healthy laying hens with eggs produced by inactivated vaccines of H5 subtype of avian influenza as the antibody.

[0042] C. In the ELISA square titration test, the purified HA protein is used as the antigen, IgY is the antibody, and biphenyldiammonium (OPD) is the substrate at 490nm, the absorbance value is 0.4 as the best ratio, and the antigen is diluted 20 The doubled concentration is 1.38mg / ml, and the antibody is...

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Abstract

The invention provides an antigen-antibody complex for treating and / or preventing avian influenza. The antigen-antibody complex contains inactivated whole avian influenza virus and its antibody as an antigen, and the mass concentration ratio of the antigen to the antibody is greater than 1. The antigen-antibody complex enters the body for initial immunization and then stimulates the body again to cause an immune response. The immune response is fast and strong; as a carrier, the antigen-antibody complex is more conducive to the capture and presentation of antigen-presenting cells, and can effectively Enhance the proliferative response of T cells; the purified whole virus as an antigen increases the molecular weight of the complex, which is more conducive to the uptake of the antigen by the immune cells of the body, and quickly causes a wider immune response. meaningful. The preparation of the antigen-antibody complex does not need to use a solid carrier, will not cause strong stimulation to the body or cause adverse reactions to the body, is simple to make, and is safe and convenient to use.

Description

Technical field [0001] The invention belongs to the field of immune regulation, and relates to an antigen-antibody complex, a preparation containing the antigen-antibody complex and a preparation method thereof, and also relates to their use in preparing medicines for preventing and / or treating avian influenza. Background technique [0002] The avian flu epidemic has caused huge losses to the world, and currently the main preventive measures are taken against it. In order to fully grasp the occurrence of the avian flu epidemic, the Chinese government also uses satellites to monitor the migration routes of migratory birds. The avian influenza vaccines developed around the world are roughly divided into inactivated influenza virus vaccines, genetic engineering subunit vaccines, DNA vaccines, recombinant liveinfluenza virus vaccines, and influenza vaccines. The genetically engineered live influenza virus vaccines and replication-deficient influenza virus vaccines are mostly inactiva...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/395A61P31/16A61K39/145
Inventor 何宏轩孙瑞芹吴艳云王承民王新卫
Owner INST OF ZOOLOGY CHINESE ACAD OF SCI
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