Vaccine composition and preparation method and application thereof

A vaccine composition and the technology of the composition are applied in the direction of pharmaceutical formulations, medical preparations containing active ingredients, antibody medical ingredients, etc., and can solve problems such as occasional allergies and transient fever

Inactive Publication Date: 2015-04-22
PU LIKE BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Chinese patent application CN103923884A discloses a porcine pseudorabies virus gene deletion strain, vaccine composition and its preparation method and application. It discloses porcine pseudorabies HN1201 strain (CCTCC NO.V201311) with deletion of gE gene, and its challenge can provide 100% protection , but there is still a transient fever for 2-3 days after the attack
[0005] The current commercialized porcine pseudorabies inactivated vaccine also has occasional allergies (Liu Yuxin et al., Diagnosis and Treatment of Piglet Pseudorabies Immune Anaphylaxis, Henan Animal Husbandry and Veterinary Medicine, 2002 (23) 2)

Method used

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  • Vaccine composition and preparation method and application thereof
  • Vaccine composition and preparation method and application thereof
  • Vaccine composition and preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0077] Example 1 Preparation of porcine pseudorabies antigen

[0078] 1.1 Preparation of Porcine Pseudorabies Antigen by Spinner Bottle Preparation Process

[0079] The porcine pseudorabies virus gE deletion strain was inoculated into PK-15 cell culture to form a seed batch, and then 1% (V / V) of the virus medium was inoculated into the monolayer PK cell culture, and rotated at 37°C Culture, when the lesion reaches 80%, harvest the toxic cell culture medium, after 2 freeze-thaw, collect the poison, and determine the poison value as 10 8.7 TCID 50 / ml. Add 10% (v / v) formaldehyde solution to the virus solution to make the final concentration of formaldehyde 0.2% (V / V), inactivate at 37°C for 18 hours, and stir every 4 hours for 10 min each time.

[0080] 1.2 Preparation of porcine pseudorabies antigen by suspension culture

[0081] The BHK-21 passaged cell line cryopreserved in liquid nitrogen has a bottom area of ​​75cm 2 The static culture in the square flask of BHK-21 sus...

Embodiment 2

[0082] Example 2 Purification of porcine pseudorabies virus

[0083] 2.1 Preparation of porcine pseudorabies purified antigen: Add 100 ml of the inactivated porcine pseudorabies antigen prepared in Example 1.2 to Triton X114 in a volume ratio of 1.0% (volume ratio) and mix well; put it at 0°C-5°C and mix for 60 minutes , and centrifuged at 37°C + 0.5°C to separate Triton X114 and virus particles. The supernatant is the purified pseudorabies virus antigen. The purified antigen of porcine pseudorabies virus is used as a standard curve with bovine serum albumin. The purified protein content is 53 μg / ml by calculating the protein content at wavelengths of 280 nm and 260 nm.

[0084] 2.2 Preparation of porcine pseudorabies purified antigen: add 100 ml of the inactivated culture of porcine pseudorabies prepared in 1.2 to NP-40 with a volume ratio of 2.0% (volume ratio) and mix well; NP-40 and virus particles were isolated by centrifugation at 37°C + 0.5°C. The supernatant was take...

Embodiment 3

[0088] Example 3 Preparation in porcine pseudorabies vaccine

[0089] Vaccine 1: The porcine pseudorabies virus inactivated antigen prepared in Example 1.1 was mixed with 206 adjuvant (product of French SEPPIC company) in a volume ratio of 54:46, and stirred at 120 rpm for 15 minutes at 30°C.

[0090] Vaccine 2: The purified porcine pseudorabies virus antigen prepared in Example 2.1 was diluted with PBS with a pH of 7.2 to a final vaccine with a protein concentration of 10 μg / head and 206 adjuvant (product of French SEPPIC) according to the volume ratio of 54:46 Mix and stir at 120 rpm for 15 minutes at 30°C.

[0091]Vaccine 3: The purified porcine pseudorabies virus antigen prepared in Example 2.1 was diluted with PBS with a P pH of 7.2 to the final vaccine protein concentration of 10 μg / head. Quil A solution was added to make the final concentration of Quil A 0.1 mg / head. Add cholesterol solution to make the final concentration of cholesterol 0.2mg / head portion, lecithin 0....

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Abstract

The invention provides a method for preparing a vaccine composition. The method comprises the following steps: (1) preparing a liquid containing a porcine pseudorabies virus; (2) adding a nonionic surfactant or a solution containing the nonionic surfactant into the liquid in the step (1) and solubilizing envelope proteins in the liquid containing the porcine pseudorabies virus; (3) removing the nonionic surfactant in the step (2) to obtain a purified porcine pseudorabies virus antigen; and (4) preparing the vaccine composition by using the antigen prepared in the step (3). The invention further provides the vaccine composition containing porcine pseudorabies antigen prepared by the preparation method. Immunosuppression components are removed from the vaccine composition and meanwhile, immunity active ingredients are retained to the maximum extent. The vaccine composition has a relatively good protective effect on porcine lung diseases; when the vaccine composition and a live virus antigen of porcine reproductive and respiratory syndrome are jointly acted, the vaccine composition has no inhibiting effect to the live virus antigen, showing that the vaccine composition can be jointly used with various other antigen components.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, in particular to a vaccine composition containing porcine pseudorabies virus antigen and a preparation method thereof. Background technique [0002] Pseudorabies, also known as Aujeszky's disease, is caused by swine herpesvirus type I (Suid herpesvirus 1strain) in the alpha subfamily of the Herpesviridae family (Herpesviridae). An acute infectious disease characterized by fever, itching (except in pigs) and encephalomyelitis. Pseudorabies in pigs is widespread in my country and is a serious disease that restricts the production of large-scale pig farms. It can cause abortion, stillbirth or mummified fetuses in pregnant sows and neurological symptoms, paralysis and high mortality in piglets. PRV has strong pantropic, neurotropic and latent infection characteristics, and can be latently infected for a long time in the peripheral nervous system. When the latent virus is acti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/245A61P31/22
Inventor 田克恭孙进忠张许科
Owner PU LIKE BIO ENG
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