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Antigen-antibody complex for preventing and/or treating avian influenza

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

Active Publication Date: 2010-06-16
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 avian influenza
  • Antigen-antibody complex for preventing and/or treating avian influenza

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0026] Concrete preparation method operates according to the following steps:

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

[0028] (2) Purified avian influenza virus from blood (healthy chicken erythrocytes treated with formaldehyde-containing physiological saline), the concentration of the antigen diluted 20 times was 1.38 mg / ml, and the concentration of the antibody diluted 20 times was 1.22 mg / ml. The antibody is the IgY prepared after multiple times of immunizing healthy laying hens with a conventional recombinant avian influenza virus H5 subtype bivalent inactivated vaccine (H5N1, Re-1 strain+Re-4 strain);

[0029] (3) Dilute the antigen at the above concentration 100 times, and dilute the...

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: formaldehyde inactivated, the whole virus of the avian influenza H5N1 virus strain purified by formaldehyde saline is the antigen.

[0034] B. Antibody: Multivalent antibody IgY prepared from eggs produced by immunizing healthy laying hens with avian influenza H5 subtype inactivated vaccine several times as an antibody.

[0035] C. In the ELISA square array titration test, take the purified influenza virus as the antigen, IgY as the antibody, biphenyl diammonium (OPD) as the substrate and read at 490nm, take the absorbance value of 0.4 as the best ratio, and dilute the antigen The concentration after 20 times is 1.38mg / ml, and the concentration after diluting the antibody 20 times is 1.22mg / ml, which is used for the preparation of antigen-antibody complexes. Dilute the antigen at the above concentration 10...

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: Extract RNA from chicken embryo allantoic fluid of avian influenza virus, perform RT-PCR, recover HA target gene, connect to pET-21b vector, transfer to BL21 competent cells for cloning and expression. The cells were lysed by ultrasonic, and the HA protein was detected by SDS-PAGE electrophoresis. Proteins were purified on a nickel column.

[0041] B. Antibody: Multivalent antibody IgY prepared from eggs produced by immunizing healthy laying hens with avian influenza H5 subtype inactivated vaccine several times as an antibody.

[0042] C. In the ELISA square matrix titration test, with the purified HA protein as the antigen, IgY as the antibody, biphenyl diammonium (OPD) as the substrate at 490nm, the absorbance value is 0.4 as the best ratio, and the antigen is diluted 20 The concentration after doubling is 1.38...

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Abstract

The invention provides an antigen-antibody complex for preventing and / or treating avian influenza, which comprises inactivation avian influenza totiviruses which are used as antigen and an immune body thereof, and the mass concentration ratio of the antigen and the immune body is more than 1. After entering organisms to perform initial immunization, the antigen-antibody complex stimulates the organisms again to induce immunoreaction, and immune response is quick in speed and strong in reaction; the antigen-antibody complex used as a carrier is more favorable for capturing and presenting antigen presenting cells and can also strengthen a breeder reaction of T cells effectively; purified totiviruses used as the antigen increase the molecular weight of the complex, are more favorable for ingestion of immunocyte of the organisms on the antigen, cause the more extensive immunoreaction quickly, and have a significance for preventing avian influenza viruses of which the antigen is easy for variation. A preparation of the antigen-antibody complex does not need to use solid carriers, does not cause intense stimulus on the organisms or initiates the organisms to generate an adverse reaction, and is simple to prepare and 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 bird flu. Background technique [0002] The epidemic of bird flu has caused huge losses to the whole world, and at present the main measures are to take preventive measures. In order to fully grasp the occurrence of bird flu, 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 whole virus inactivated vaccines (inactivated influenza virus vaccines), genetically engineered subunit vaccines (subunit vaccines), DNA vaccines (DNA vaccines), recombinant live vector vaccines (recombine liveinfluenza virus vaccines), influenza Genetically engineered live influenza virus...

Claims

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Application Information

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Patent Type & Authority Applications(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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