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Preparation method of porcine parvovirus inactivated vaccine

A parvovirus and inactivated vaccine technology, applied in the field of bioengineering, can solve the problem of low antibody titer, achieve the effect of increasing antigen content, extensive social and economic benefits, and improving immune effect

Inactive Publication Date: 2012-09-26
扬州优邦生物药品有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Immune adjuvant is also an important factor affecting the immune effect of porcine parvovirus inactivated vaccine, but the mixture of several adjuvants, such as oil adjuvant, SDS, L-121, mixture of aluminum hydroxide and oil emulsion and SDS, aluminum hydroxide The antibody titers produced by vaccines adjuvanted with the mixture of

Method used

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  • Preparation method of porcine parvovirus inactivated vaccine
  • Preparation method of porcine parvovirus inactivated vaccine
  • Preparation method of porcine parvovirus inactivated vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] This implementation case illustrates the method for porcine parvovirus venom preparation:

[0027] 1. Cell preparation Take the cells out of the liquid nitrogen tank and place them in a 37°C water bath to melt them quickly. Transfer the cells into a cell bottle containing 10% fetal bovine serum cell growth medium and culture them at 37°C. When they grow into a good monolayer, use pancreatic Digest the cells with enzymatic digestion solution, and expand step by step according to 1:2 passaging. Inoculate the expanded seed cells into 15000ml spinner bottles, add 1000ml of cell growth solution containing 10% newborn bovine serum to each bottle, culture at 37°C, spin the bottle at a speed of 6-12 rpm, and grow into a good monolayer Afterwards, the cells were digested with trypsin solution, and divided into bottles at a ratio of 1:3 for passage.

[0028] 2. Virus propagation Take F13 generation BJ-2 strain PPV to produce seed virus, inoculate well-grown ST cells at 2% inocul...

Embodiment 2

[0031] This implementation case illustrates the method of purifying and concentrating porcine parvovirus:

[0032] Take 20,000 milliliters of porcine parvovirus liquid with a hemagglutination value of 28, purify, concentrate, measure the hemagglutination value, and compare the difference before and after concentration. Specific steps are as follows:

[0033] (1) Add the raw material solution to a microfiltration system with a pore size of 0.22 microns for filtration, return the concentrated solution to the raw material tank, and discharge the raw material solution after about 10 minutes; collect the osmotic pressure, and observe the permeate at this time, it is clear and bright, and it has been removed Impurities such as cell debris enter the next step of ultrafiltration concentration.

[0034] (2) Ultrafiltration concentration of parvovirus venom

[0035] Concentrate the pretreated permeate with a 1m2 polyethersulfone flat membrane bag with a molecular weight cut-off of 30K...

Embodiment 3

[0050] This implementation case illustrates the method of porcine parvovirus inactivation:

[0051] Diethyleneimine (BEI) inactivation test Take 5 bottles of porcine parvovirus antigen solution, add 0.02%, 0.05%, 0.08%, 0.1%, 0.15% final concentration of BEI (made by BEA cyclization), 32 ℃ Inactivation, samples were taken at 24, 48, 72, 96, and 120 hours of inactivation for inactivation testing, and the inactivated venom samples were diluted 10 times with serum-free cell nutrient solution, and then inoculated with cells that had grown into a single layer. Porcine testicular cells (ST cells), 1ml / bottle, after 1 hour of adsorption, replace with cell growth medium containing 2% fetal bovine serum and culture for 5 days, observe cell morphology and growth status, and determine whether there is cytopathic disease (CPE), The culture was harvested, frozen and thawed 3 times, and continuously passed on ST cells for 2 generations. At the same time, HA was detected to observe whether t...

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Abstract

The invention relates to a preparation method of a porcine parvovirus inactivated vaccine. The preparation method comprises the following preparation steps of: (1) taking a porcine parvovirus fluid out of a raw material tank, placing the porcine parvovirus fluid into a microfiltration system, filtering to obtain a penetrating fluid and a concentrated solution, returning the concentrated solution to the raw material tank, and discharging the porcine parvovirus fluid after the solid content of the porcine parvovirus fluid is larger than or equal to 1g / L; (2) carrying out ultrafiltration treatment on the penetrating fluid, and further concentrating the porcine parvovirus fluid until the virus hemocoagulate value is up to 28-213 to obtain a concentrated solution; (3) filtering the concentrated solution again by using the microfiltration system to obtain a parvovirus fluid with the hemocoagulate value of 28-213 after the treatment; and (4) inactivating the parvovirus fluid and preparing the concentrated and inactivated parvovirus fluid into a vaccine product. The invention provides a safe and feasible vaccine production method so that vaccine production is not dependent on the traditional process again, high-titer parvovirus fluid is produced, the antigen content is increased, and the requirement for immune production is met.

Description

technical field [0001] The invention relates to a preparation method of an inactivated porcine parvovirus vaccine, in particular to a method for designing a purification and concentration process of an inactivated porcine parvovirus vaccine, belonging to the technical field of bioengineering. Background technique [0002] Porcine parvovirus (PPV) can cause reproductive disorders in sows, resulting in stillbirths, mummified fetuses, early embryonic death and infertility in sows. At present, the disease has been distributed worldwide and has caused serious economic losses to the pig industry. German scholar Mayr first discovered PPV for the first time, and Chinese scholar Pan Xuezhu and others successfully isolated the virus in 1983. [0003] At present, PPV vaccines mainly include inactivated vaccines, attenuated vaccines, genetically engineered subunit vaccines, and genetically engineered live virus vector vaccines. Strong efficacy, fast antibody production, less dosage, l...

Claims

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Application Information

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IPC IPC(8): A61K39/23A61P31/20
Inventor 潘杰范娟沈明君傅元华钱钟
Owner 扬州优邦生物药品有限公司
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