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Preparation method of inactivated lyophilized vaccine for porcine circovirus type 2

A porcine circovirus, inactivation technology, applied in freeze-drying delivery, antiviral agents, viral antigen components, etc., can solve the problems of poor immune effect, high side reaction rate, difficult to absorb, etc., and achieve easy injection and high antigen content. , the effect of easy absorption

Inactive Publication Date: 2015-03-04
TIANJIN RINGPU BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention aims at the technical defects of the prior art, and provides a preparation method of porcine circovirus type 2 inactivated freeze-dried vaccine to solve the technical problems of high side reaction rate, poor immune effect, difficult injection and difficult absorption

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1 (vaccine preparation method)

[0031] 1 Instruments and reagents

[0032] 1.1 The instrument used in the embodiment is as follows:

[0033] Milipore’s 0.5-0.2um double-layer filter element; Cobetter sleeve; Milipore’s cross-cut UF concentration membrane package, 100KD; GE’s XK50 / 100 column; GE’s AKTA Primer plus chromatography system; Bio-Rad Protein detector; Pudong small freeze dryer.

[0034] 1.2 The reagents used in the embodiment are as follows:

[0035] (1) 100L porcine circovirus type 2 virus liquid, 10 7.2 TCID 50 / ml, produced by our company;

[0036] (2) 100L of 1M NaOH; 100L of 0.1M PBS; 100L of high-purity water;

[0037] (3) Trehalose, sucrose, lactose, skim milk, glycine.

[0038] 2 operation steps

[0039] 2.1 Clarification of porcine circovirus type 2 venom

[0040] Under sterile conditions, the harvested venom is filtered through 8 layers of sterile gauze, and the primary filtered venom is clarified through a 0.5um-2um double-layer...

Embodiment 2

[0079] Embodiment 2 (vaccine safety test prepared in embodiment 1)

[0080] The PCV2 inactivated freeze-dried vaccine 1 prepared by the above process, the PCV2 white oil adjuvant inactivated vaccine 2 prepared by the conventional process, and sterile 0.01M PBS solution were used as experimental reagents; 20 days old PCV2 ELISA antibody negative, PCV2 antigen negative healthy easy 15 affected piglets were used as experimental animals. The specific operation steps are as follows:

[0081] 1 animal group

[0082] 15 piglets were randomly divided into 3 groups, 5 piglets in each group.

[0083] 2 vaccine immunization

[0084] Inject the two vaccines and PBS into the experimental piglets intramuscularly in the neck, as follows:

[0085] In the first group, 4ml PCV2 inactivated freeze-dried vaccine 1 was injected intramuscularly into the neck of each piglet, a total of 5 piglets;

[0086] The second group, the PCV2 white oil adjuvant inactivated vaccine 2 that every head piglet...

Embodiment 3

[0095] Embodiment 3 (the vaccine potency test that embodiment 1 prepares)

[0096] 1 Mice challenge test

[0097] PCV2 inactivated freeze-dried vaccine 1, PCV2 white oil adjuvanted inactivated vaccine 2 prepared by conventional techniques, and sterile 0.01M PBS solution were used as experimental reagents; 30 6-week-old healthy Balb / C mice were used as experimental animals. The specific experimental steps are as follows:

[0098] 1.1 Animal grouping

[0099] Thirty Balb / C mice were randomly divided into 3 groups, 10 in each group.

[0100] 1.2 Vaccine Immunization

[0101] Two kinds of vaccines and PBS were intraperitoneally injected into experimental mice, as follows:

[0102] In the first group, each mouse was intraperitoneally injected with 0.2ml PCV2 inactivated freeze-dried vaccine 1, a total of 10 mice;

[0103] The second group, each mouse was intraperitoneally injected with 0.2ml PCV2 white oil adjuvant inactivated vaccine 2 prepared by conventional technology, tot...

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Abstract

The invention provides a preparation method of an inactivated lyophilized vaccine for porcine circovirus type 2. The preparation method comprises the following steps: successively carrying out clarification procedure, ultrafiltration concentration and molecular sieve chromatography column purification on a virus culture solution, so that the impurity content of an antigen used for preparing the vaccine is effectively ensured to be reduced to the minimum, thus avoiding the phenomenon that side effects are generated after the product is used, and also being beneficial to ensuring the immune effect of the vaccine. In addition, according to the preparation method provided by the invention, as the inactivated antigen is directly prepared in a lyophilizing mode without adding an oil adjuvant, the problems of difficulty in injection and absorption do not exist. The PCV 2 inactivated lyophilized vaccine prepared by the preparation method is high in antigen content, immunogenicity and antigen purity, is good in safety, is low in side effect, is capable of rapidly generating antibody protection without an adjuvant and is easily injected and absorbed.

Description

technical field [0001] The invention relates to the technical field of vaccine preparation, in particular to a preparation method of porcine circovirus type 2 inactivated freeze-dried vaccine. Background technique [0002] Porcine circovirus disease is a series of diseases caused by porcine circovirus type 2 (PCV2), including postweaning multisystemie wasting syndrome (PMWS), porcine dermatitis and nephrotic syndrome ( Porcine dermatitis and nephropathy syndrome, PDNS), porcine respiratory disease complex (Porcine respiratory disease complex, PRDC). [0003] In 1997, Clark et al. isolated porcine circovirus type 2 for the first time. At the 20th International Swine Disease Conference in 2008, circovirus disease was listed as the number one disease currently endangering the development of the pig industry. In 2000, my country confirmed the existence of PCV2 infection in pig herds. In 2002, PWMS caused by PCV2 infection was an outbreak in the whole country. Porcine circovir...

Claims

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Application Information

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IPC IPC(8): A61K39/12A61K9/19A61K47/42A61P31/20
Inventor 唐荣宏吕茂杰李亚杰杨保收梁武
Owner TIANJIN RINGPU BIO TECH
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