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 ad

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

Example Embodiment

[0025] Example 1

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

[0027] 1. Cell preparation. Remove the cells from the liquid nitrogen tank and place them in a 37℃ water bath to quickly melt them. Transfer the cells into a cell bottle containing 10% fetal bovine serum cell growth solution and culture at 37℃. Use pancreas when it grows into a good monolayer. Enzyme digestion solution digests the cells, and gradually expands by 1:2 passage. Inoculate the expanded seed cells into 15000ml spinner flasks, add 1000ml cell growth medium containing 10% newborn calf serum to each bottle, culture at 37℃, spinner flask speed 6-12 revolutions / hour, wait to grow into a good monolayer Afterwards, the cells were digested with trypsin digestion solution and passaged in 1:3 flasks.

[0028] 2. Virus propagation Take F13 generation BJ-2 strain PPV to produce seed virus, inoculate well-growing ST cells at 2% inoculation amount, and add cell maintenance solution co...

Example Embodiment

[0030] Example 2

[0031] This case illustrates the method of purification and concentration of porcine parvovirus:

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

[0033] (1) Add the raw material liquid to a microfiltration system with a pore size of 0.22 microns for filtration. The concentrated liquid returns to the raw material tank. After about 10 minutes, the raw material liquid is discharged; collect the osmotic pressure, and observe that the permeate is clear and bright. Cell debris and other impurities enter the next step of ultrafiltration and concentration.

[0034] (2) Ultrafiltration concentration of parvovirus venom

[0035] The pretreated permeate was concentrated with a 1m2 polyethersulfone flat membrane package with a molecular weight cut-off of 30KD to further concentrate the venom. The c...

Example Embodiment

[0049] Example 3

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

[0051] Diethyleneimine (BEI) inactivation test Take 5 bottles of porcine parvovirus antigen solution and add 0.02%, 0.05%, 0.08%, 0.1%, 0.15% final concentration of BEI (made by BEA cyclization), 32℃ Inactivate, take samples at 24, 48, 72, 96, and 120 hours of inactivation for inactivation test. Dilute the inactivated venom sample 10 times with serum-free cell nutrient solution, and then inoculate the monolayer Porcine testicular cells (ST cells), 1ml / bottle, after adsorption for 1 hour, change to cell growth medium containing 2% fetal bovine serum and culture for 5 days, observe the cell morphology and growth status, and determine whether there is cytopathic (CPE). Harvest the culture, freeze and thaw 3 times, pass it on ST cells for 2 consecutive generations, and do HA test at the same time to observe whether there is agglutination. The test results are now reported as...

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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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