Preparation method of porcine circovirus type 2 inactivated vaccine

A porcine circovirus and inactivated vaccine technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, antiviral agents, etc., can solve the problems of low safety and large side effects, and achieve high safety, small side effects, The effect of high product quality controllability

Active Publication Date: 2014-03-12
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

[0004] The currently used PCV-2 vaccines all use mineral oil as an adjuvant,

Method used

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  • Preparation method of porcine circovirus type 2 inactivated vaccine
  • Preparation method of porcine circovirus type 2 inactivated vaccine
  • Preparation method of porcine circovirus type 2 inactivated vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] This example illustrates the determination of PK-15 cell culture circovirus bioreactor culture conditions and PCV-2 proliferation conditions.

[0027] (1) Put 2×10 5 PK-15 cells per ml were inserted into the reactor, supplemented with DMEM medium containing 5% bovine serum to 5L, set the stirring speed at 90 rpm, dissolved oxygen at 50%, pH 7.2, and temperature 37 ℃, carry out cell suspension culture, turn on the inlet pump and outlet pump for fed-batch culture after 24 hours, take regular samples every day to detect the consumption of glucose and record the consumption of lye (1%NaOH), and continue to cultivate for 13 days. Open the jar, digest and count the cells. The final culture conditions were determined as follows: 90 rpm, dissolved oxygen 50%, pH 7.2, temperature 37°C.

[0028] (2) When the cell reactor is cultivated for 48 hours, 72 hours and 96 hours, the cell growth liquid is discharged, the maintenance liquid containing 1% serum is inserted into the reacto...

Embodiment 2

[0030] This example illustrates the method of inoculating and culturing the virus to prepare the virus solution.

[0031] The volume is 1L and the concentration is 2~3×10 5 PK-15 cells per ml were inserted into the reactor, supplemented with DMEM medium containing 5% bovine serum to 5L, set the stirring speed at 90 rpm, dissolved oxygen at 50%, pH 7.2, and temperature 37 ℃, carry out cell suspension culture, after 24 hours, turn on the liquid inlet pump and the liquid outlet pump for fed-batch culture; after the cell reactor is cultivated for 72 hours, the cell growth liquid is discharged, and the maintenance liquid containing 1% serum is inserted into the reactor , insert the prepared circovirus (YZ strain) F15 seed virus, the inoculation volume is 400mL, add D-glucosamine with a final concentration of 2mmol / L to the virus maintenance solution, adjust the pH to 7.4, and set the temperature At 36.5°C, harvest the venom after 24 hours of cultivation and replenish fresh mainten...

Embodiment 3

[0033] This example illustrates the method of inactivating virus liquid and preparing vaccine products.

[0034] Inactivation of virus fluid:

[0035]The virus liquid finally obtained in Example 2 was collected and inactivated with cyclized BEI with a final concentration of 1.0‰ at a central temperature of 32°C for 48 hours, and then terminated with sterile sodium thiosulfate solution with a final concentration of 2%.

[0036] Preparation of inactivated vaccine:

[0037] Take 1 part of MontanideTM ISA206VG adjuvant from Sepic Company in France, 1 part of qualified inactivated antigen, mix them in an equal amount in an emulsification tank, stir and emulsify at a low speed for 30 minutes, and prepare PCV-2 (YZ strain) water-in-oil-in-water Dosage forms of inactivated vaccines.

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Abstract

The invention relates to a preparation method and a product of a porcine circovirus type 2 inactivated vaccine, and belongs to the technical field of biological products. The preparation method of the porcine circovirus II-type inactivated vaccine disclosed by the invention comprises the steps of 1) implementing multiplication culture on a PK-15 cell; 2) inoculating virus to culture and prepare a virus solution; and 3) condensing and inactivating the virus, and preparing a finished vaccine product. The preparation method disclosed by the invention, which amplifies PCV-2 (porcine circovirus-2) through a continuous perfusion-type bioreactor, has the advantages of being relatively small in operation space, simple in operation process, remarkably low in labor, capable of being used for mass production through cascade amplification, and being low in serum content in maintenance media, relatively low in production cost and high in product quality controllability; the prepared porcine circovirus type 2 inactivated vaccine has the advantages of high safety, good immune effect, low side effect and the like.

Description

technical field [0001] The invention relates to a preparation method and product of porcine circovirus type 2 inactivated vaccine, belonging to the technical field of biological products. Background technique [0002] Porcine circovirus (PCV) first broke out in Canada in 1997, and subsequently reported the emergence of the disease in many countries around the world. PCV has two genotypes, PCV1 and PCV-2. Tischer et al. first discovered the existence of PCV-1 in PK-15 cells. %about. Pig kidney epithelial cells PK-15 are derived from pig kidneys. The Chinese name is pig kidney epithelial cells. The normal cell morphology is epithelioid and grows adherently. The cells are sensitive to a variety of viruses, such as porcine circovirus (PCV), porcine parvovirus (PPV), swine fever virus (CSFV), etc., and can be applied to porcine circovirus vaccine, porcine parvovirus vaccine, and swine fever virus Preparation of vaccines, etc. PCV1 does not cause disease in pigs, but PCV-2 can...

Claims

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Application Information

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IPC IPC(8): A61K39/12A61P31/20C12N7/00C12R1/93
Inventor 范娟李群刘俊斌钱钟宋庆庆
Owner 扬州优邦生物药品有限公司
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