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Cell inactivated vaccine, egg yoke antibody and injection and freeze-dried powder containing same

An inactivated vaccine and egg yolk antibody technology, which is applied in the direction of antibody, freeze-dried delivery, powder delivery, etc., can solve the problems of cumbersome preparation methods of egg yolk antibody, single treatment, antibody loss, etc., achieve good clinical application effect and enhance immunogenicity , the effect of reducing the loss of antibodies

Inactive Publication Date: 2013-01-09
河南后羿生物工程股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, the treatment of anti-porcine circovirus type 2 vaccine is relatively simple, and the preparation method of egg yolk antibody is cumbersome, and the loss of antibody is relatively high.

Method used

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  • Cell inactivated vaccine, egg yoke antibody and injection and freeze-dried powder containing same
  • Cell inactivated vaccine, egg yoke antibody and injection and freeze-dried powder containing same
  • Cell inactivated vaccine, egg yoke antibody and injection and freeze-dried powder containing same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The cell inactivated vaccine provided in this embodiment is prepared by the following method:

[0034] 1) Take the disease material to make a homogenate, freeze and thaw repeatedly 3 times, centrifuge at 10000-12000rpm for 15-20min, take the supernatant, filter it through a 0.22μm filter membrane, and inoculate a sensitive monolayer of PK-15 cells;

[0035] 2) The PK-15 cells were continuously passaged, the cells were frozen and thawed, and the DNA of the PCV2 virus was extracted. Aiming at the ORF1 gene of the PCV2 virus, the nucleotide sequences of SEQ ID NO: 1 and SEQ ID NO: 2 in the sequence table and SEQ ID Two pairs of specific PCR primers composed of the nucleotide sequences of NO: 3 and SEQ ID NO: 4 amplify the target gene, wherein,

[0036] Primer 1PCV2.1: 5'-TTCGGTACCAGCTATGACGTATCCAAG-3' (SEQ ID NO: 1),

[0037] PCV2.2: 5'-GCCAAGCTTTCACTTCGTAATGGTTTT-3' (SEQ ID NO: 2);

[0038] Primer 2PCV2.1: 5'-TTGGCCTAAGCTATGACGTATCCAA-3' (SEQ ID NO: 3),

[0039] PCV2.2...

Embodiment 2

[0046] The yolk antibody provided in this example was prepared by the following method:

[0047] 1) Use bacterial inactivated vaccines to carry out basic immunization, booster immunization and booster immunization for healthy laying hens every 7 days. The basic immunization vaccination is 1.5ml, the dose of booster immunization and booster immunization is doubled, and the booster immunization Then start to detect the antibody titer. When the antibody titer of porcine circovirus reaches 1:64 and goose circovirus antibody titer reaches 1:64 or more, collect high-immune eggs;

[0048] 2) Clean under running water, sterilize the high-free eggs with 84 disinfectant solution, dry them in the air, take the egg yolk by aseptic technique, and obtain the egg yolk stock solution;

[0049] 3) Add an equal volume of sterile water to the egg yolk stock solution, stir and mix until the color turns white, and obtain the egg yolk solution;

[0050] 4) Put the egg yolk solution in a 60°C water...

Embodiment 3

[0054] The yolk antibody provided in this example was prepared by the following method:

[0055] 1) Use bacterial inactivated vaccines to carry out basic immunization, booster immunization and booster immunization on healthy laying hens every 10 days. The basic immunization vaccine is 1.5ml, the booster immunization and booster immunization are doubled, and the booster immunization Then start to detect the antibody titer. When the antibody titer of porcine circovirus reaches 1:64 and goose circovirus antibody titer reaches 1:64 or more, collect high-immune eggs;

[0056] 2) Clean the eggs with running water, sterilize the high-free eggs with bromogeramine, dry them in the air, take the egg yolk by aseptic technique, and obtain the egg yolk stock solution;

[0057] 3) Add an equal volume of sterile water to the egg yolk stock solution, stir and mix until the color turns white, and obtain the egg yolk solution;

[0058] 4) Put the egg yolk solution in a 65°C water bath and heat...

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Abstract

The invention discloses cell inactivated vaccine which simultaneously contains porcine circovirus epitopes and goose circovirus epitopes. The cell inactivated vaccine is prepared by the method including screening high-pathogenicity strains from porcine circoviruses; selecting a plurality of virus epitopes; connecting the epitopes with one another; then inserting the connected epitopes into genomes of goose circoviruses to proliferate; and adding Freund adjuvants to formaldehyde for inactivation so as to obtain the cell inactivated vaccine. The invention simultaneously discloses an egg yolk antibody prepared from the cell inactivated vaccine, injection and freeze-dried powder containing the egg yolk antibody. Genes of the porcine circovirus are inserted into the genomes of the goose circoviruses, the immunogenicity of the goose circoviruses is strengthened, immune systems of goose bodies are excited, so that the titer of generated antibodies is higher than that of egg yolk antibodies generated by porcine circovirus vaccine used singularly, a clinical application effect is good, and generated circovirus antibodies have an auxiliary treatment effect for porcine circovirus diseases.

Description

technical field [0001] The invention relates to a cell inactivated vaccine, egg yolk antibody and injection and freeze-dried powder of egg yolk antibody, belonging to the field of disease immunity. Background technique [0002] Porcine circovirus type 2 (PVC2) is the main pathogen of multisystem failure syndrome and nephrotic dermatitis syndrome in weaned piglets. At the same time, the disease can also cause immunosuppression, which is easy to cause secondary or concurrent infection of other diseases, with a high incidence , high prevalence, high pathogenicity, high mortality and other characteristics have caused huge economic losses to the pig industry. The pathogenic mechanism of PVC2 is still unclear, and its destruction of the pig immune system is considered to be the main cause of related pig diseases. Due to the decline of immunity, pigs are vulnerable to other pathogens during stress and pathogenic infection. The main clinical symptoms are progressive emaciation, ane...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/12A61K39/42A61K9/08A61K9/19A61P31/20C12N7/01C07K16/08C07K16/02C12R1/93
Inventor 吴红云郭俊清徐进
Owner 河南后羿生物工程股份有限公司
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