Vaccine compound 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 transient fever and occasional allergies

Inactive Publication Date: 2019-04-05
PULIKE BIOLOGICAL ENG INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] 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
[0006] 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 compound and preparation method and application thereof
  • Vaccine compound and preparation method and application thereof
  • Vaccine compound and preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0078] The preparation of embodiment 1 porcine pseudorabies antigen

[0079] 1.1 Preparation of porcine pseudorabies antigen by spinner bottle preparation process

[0080] The gE-deleted strain of porcine pseudorabies virus was inoculated in PK-15 cell culture to form a seed batch, and then inserted into the PK cell culture to form a monolayer according to 1% (V / V) of the virus culture solution, and rotated at 37°C Cultivate, when the lesion reaches 80%, harvest the toxic cell culture medium, after freezing and thawing twice, collect the poison, and measure the poison price as 10 8.7 TCID 50 / ml. Add 10% (v / v) formaldehyde solution to the virus solution so that the final concentration of formaldehyde is 0.2% (V / V), inactivate at 37°C for 18 hours, stir once every 4 hours, and stir for 10 minutes each time.

[0081] 1.2 Preparation of porcine pseudorabies antigen by suspension culture method

[0082] The bottom area of ​​the BHK-21 passage cell line frozen in liquid nitroge...

Embodiment 2

[0083] Embodiment 2 purifies porcine pseudorabies virus

[0084] 2.1 Preparation of the purified antigen of porcine pseudorabies: add 100ml of the inactivated antigen of porcine pseudorabies prepared in Example 1.2 into Triton X114 with a volume ratio of 1.0% (volume ratio) and mix; put it at 0°C-5°C and mix for 60 minutes , placed at 37°C+0.5°C and centrifuged to separate Triton X114 and virus particles. The supernatant is the purified pseudorabies virus antigen. The purified porcine pseudorabies virus antigen is used as a standard curve with bovine serum albumin, and the protein content is calculated at 280nm and 260nm wavelengths. The purified protein content is 53 μg / ml.

[0085] 2.2 Preparation of the purified antigen of porcine pseudorabies: add 100 ml of the inactivated culture of porcine pseudorabies prepared in 1.2 into NP-40 with a volume ratio of 2.0% (volume ratio) and mix; put it at 0°C-5°C and mix for 60 minutes, Centrifuge at 37°C+0.5°C to separate NP-40 and vi...

Embodiment 3

[0089] Preparation in embodiment 3 porcine pseudorabies vaccine

[0090] Vaccine 1: Mix the porcine pseudorabies virus inactivated antigen prepared in Example 1.1 with 206 adjuvant (product of SEPPIC, France) at a volume ratio of 54:46, and stir at 120 rpm for 15 minutes at 30°C.

[0091] Vaccine 2: The purified porcine pseudorabies virus antigen prepared in Example 2.1 is diluted with PBS with a pH of 7.2 to a final vaccine protein concentration of 10 μg / head portion and 206 adjuvant (French SEPPIC company product) according to the volume ratio of 54:46 Mix and stir at 120 rpm for 15 minutes at 30°C.

[0092]Vaccine 3: The purified porcine pseudorabies virus antigen prepared by Example 2.1 is diluted with PBS with P pH of 7.2 to a protein concentration of 10 μg / head in the final vaccine. Add Quil A solution so that the final concentration of Quil A is 0.1mg / head Parts, add cholesterol solution so that the final concentration of cholesterol is 0.2mg / head portion, lecithin 0.2...

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Abstract

The invention provides a method for preparing a vaccine compound. The method comprises the steps that 1, liquid containing porcine pseudorabies virus is prepared; 2, by adding a nonionic surfactant ora solution containing the nonionic surfactant into the liquid of the step 1, envelope protein which exists in the porcine pseudorabies virus liquid is solubilized; 3, the nonionic surfactant in the step 2 is removed to obtain an purified porcine pseudorabies virus antigen; 4, the antigen prepared in the step 3 is used for preparing the vaccine compound. The invention further provides the vaccinecompound which is prepared through the preparation method and contains the porcine pseudorabies antigen; according to the vaccine compound, an immunosuppression component is removed, and meanwhile, animmunocompetence component is maintained furthest, so that the vaccine compound has a good protective effect on lung lesion of pigs; when the vaccine compound is used in cooperation with a porcine reproductive and respiratory syndrome live virus antigen, the vaccine compound does not have an inhibiting effect on the porcine reproductive and respiratory syndrome live virus antigen, which shows that the vaccine compound can be used in cooperation with multiple kinds of other antigen components.

Description

[0001] This application is a divisional application of patent application 201410763549.6. The original invention title is "vaccine composition and its preparation method and application", and the original filing date is December 12, 2014. technical field [0002] The invention belongs to the technical field of veterinary biological products, and in particular relates to a vaccine composition containing porcine pseudorabies virus antigen and a preparation method thereof. Background technique [0003] Pseudorabies, also known as Aujeszky's disease, is a disease caused by swine herpesvirus type Ⅰ (Suid herpesvirus 1strain) in the α subfamily of the Herpesviridae family (Herpesviridae) in pigs, cattle, sheep and other domestic animals, poultry and wild animals. An acute infectious disease characterized by fever, severe itching (except pigs) and encephalomyelitis. Pseudorabies in pigs is widespread in our country and is seriously harmful. It is one of the main diseases restrictin...

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 PULIKE BIOLOGICAL ENG INC
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