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Porcine circovirus type 2, mycoplasma hyopneumoniae and haemophilus parasuis triple inactivated vaccine and preparation method thereof

A technology of Mycoplasma hyopneumoniae and Haemophilus suis, which is applied in the direction of virus antigen components, virus/phage, biochemical equipment and methods, etc., can solve problems such as difficulty in guaranteeing 100% activity, lack of processing and modification, and improve pig production Good performance, good mental state, good protein activity

Inactive Publication Date: 2020-02-21
山东滨州沃华生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The Escherichia coli expression system is a prokaryotic expression system, which lacks post-translational processing and modification of the protein. Therefore, it is impossible to modify the spatial configuration of the expressed circular cap protein, and it is difficult to ensure 100% activity of the VLP of the late cap protein.

Method used

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  • Porcine circovirus type 2, mycoplasma hyopneumoniae and haemophilus parasuis triple inactivated vaccine and preparation method thereof
  • Porcine circovirus type 2, mycoplasma hyopneumoniae and haemophilus parasuis triple inactivated vaccine and preparation method thereof
  • Porcine circovirus type 2, mycoplasma hyopneumoniae and haemophilus parasuis triple inactivated vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] The preparation of embodiment 1 porcine circovirus type 2, mycoplasma hyopneumoniae, Haemophilus parasuis triple inactivated vaccine, such as Figure 5 Shown:

[0048] 1. Preparation of porcine circovirus type 2 semi-finished antigen

[0049] 1) The preferred eukaryotic expression plasmid containing circovirus type 2 cap protein was electrotransformed into Pichia pastoris cells (X-33, purchased from Invitrogen, USA). Positive clone seeds were screened out, inoculated in 250ml of BMGY liquid medium, 30°C, 250r / min shaking culture for 16h-24h; after that, 250ml of proliferated seed liquid was added to sterilized FBSM inorganic salt medium, and fermented in a fermenter Expanded cultivation in. When the OD600 of the bacterial liquid was 1.0, 1% methanol was added to induce it for 80h-96h, and the bacterial liquid was harvested.

[0050] 2) The yeast harvested liquid was centrifuged to remove the supernatant, and the precipitate was re-selected with 0.01M PBS solution and c...

Embodiment 2

[0068] Example 2 Safety Test of Porcine Circovirus Type 2, Mycoplasma Hyopneumoniae, and Haemophilus Parasuis Triple Inactivated Vaccine

[0069] 1. Guinea pig security inspection: Divide 180-220g guinea pigs into two groups, 8 in each group. In the test group, each guinea pig was intraperitoneally injected with 5ml of porcine triple vaccine; in the control group, each guinea pig was intraperitoneally injected with 5ml of normal saline, and observed continuously for 7 days. All the guinea pigs in the two groups survived without local or systemic adverse reactions. The results are shown in Table 1. .

[0070] Table 1 pig triple vaccine of the present invention is in the safety inspection result of guinea pig

[0071]

[0072]

[0073] 2. Piglet security check: screen three litters of healthy piglets for 14-21 days, select 6 piglets in each litter, inject 2ml of the pig triple vaccine of the present invention into the neck muscle, observe continuously for 2 hours, and no ...

Embodiment 3

[0074] Example 3 The protective effect of pig triple vaccine on circovirus type 2 and Haemophilus parasuis in mice

[0075] 1. The protective effect of triple vaccine against circovirus type 2 challenge

[0076] 30 18-22g BALB / C mice were fed for one week and then divided into 3 groups, namely triple vaccine group, single circular vaccine group and blank control group. Each mouse was intraperitoneally injected with 0.5ml vaccine, and the control group was injected with the same dose of PBS solution. After 14 days, booster immunization was carried out according to the same method, with a dose of 0.5ml / mouse.

[0077] Twenty-one days after BALB / c mice were immunized, together with 10 control mice, the DBN-SX07 strain was used to test the virus (≥10 7.0 TCID 50 / ml) challenge, each mouse was intraperitoneally injected with 0.5ml, and 21 days after the challenge, the spleen was culled and taken for virus isolation. If the virus was isolated, it was judged to be positive for viru...

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Abstract

The invention discloses a porcine circovirus type 2, mycoplasma hyopneumoniae and haemophilus parasuis triple inactivated vaccine and a preparation method thereof, which belong to the technical fieldof veterinary biological products. The vaccine consists of an antigen and a vaccine adjuvant, wherein the antigen consists of a porcine circovirus type 2 antigen, a mycoplasma hyopneumoniae antigen and a haemophilus parasuis antigen; the porcine circovirus type 2 antigen is cap protein obtained through expression of pichia pastoris, and the protein content is larger than or equal to 150 microgramsper milliliter, the mycoplasma hyopneumoniae antigen is inactivated mycoplasma hyopneumoniae bacterial liquid; wherein the haemophilus parasuis is an inactivated haemophilus parasuis liquid; a vaccine adjuvant is composed of a water-based high-molecular polymer adjuvant and a composite polysaccharide immunopotentiator. The triple inactivated vaccine for the porcine circovirus type 2, the mycoplasma hyopneumoniae and the haemophilus parasuis provided by the invention has obvious advantages in preventing the three swine diseases and improving the swine production performance.

Description

technical field [0001] The invention relates to the field of veterinary biological products, in particular to a triple inactivated vaccine of porcine circovirus type 2, mycoplasma hyopneumoniae and Haemophilus parasuis and a preparation method thereof. Background technique [0002] Porcine circovirus disease is an infectious disease of pigs caused by porcine circovirus type 2, which causes piglet weaning syndrome, porcine dermatitis and nephrotic syndrome, and reproductive disorders in sows. Its clinical manifestations are diverse. After pigs are infected with circovirus, it can cause severe immunosuppression, resulting in secondary or concurrent other infectious diseases, such as Mycoplasma hyopneumoniae, Haemophilus parasuis, Streptococcus suis and other diseases. [0003] Mycoplasma swine pneumonia, also known as porcine endemic pneumonia, is a respiratory disease caused by Mycoplasma hyopneumoniae, characterized by chronic, highly contagious, high morbidity and low morta...

Claims

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Application Information

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IPC IPC(8): A61K39/295A61K39/12A61K39/02A61K39/102A61P31/04A61P31/20
CPCA61K39/12A61K39/0241A61K39/102A61P31/04A61P31/20C12N2750/10034A61K2039/5252A61K2039/70A61K2039/552A61K2039/55583A61K2039/521
Inventor 耿兴良王楠朱杰徐海军杨合涛杨灵芝
Owner 山东滨州沃华生物工程有限公司
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