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A kind of porcine epidemic diarrhea virus vaccine and preparation method thereof

A porcine epidemic diarrhea virus vaccine technology, applied in the field of porcine epidemic diarrhea virus vaccine and preparation thereof, can solve the problems of difficulty in preparing porcine epidemic diarrhea virus inactivated vaccine, decreased resistance of piglets, unfavorable protection of piglets, etc. Significant difference in titer, good immune protection, and the effect of improving the level of specific SIgA

Active Publication Date: 2020-12-01
天津中逸安健生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it was found in the study that although the vaccine can produce antibodies quickly after the vaccine is injected into pregnant sows 42 days before delivery, the antibody level shows a significant downward trend in the second week after immunization, and the maternal antibody level of piglets after weaning The level gradually subsides, resulting in a decrease in the resistance of piglets, which is not conducive to the protection of piglets after weaning
In addition, the vaccine uses a variety of adjuvant components made of squalane, oleic acid, and Tween 80. The components are complex, which also brings certain difficulties to the preparation of porcine epidemic diarrhea virus inactivated vaccines.

Method used

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  • A kind of porcine epidemic diarrhea virus vaccine and preparation method thereof
  • A kind of porcine epidemic diarrhea virus vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] (1) Dissolve 10mg of PBD-b-PEO and 1mg of DOTAP in 10ml of tetrahydrofuran solution. After the DOTAP lipid is dissolved, a PBD-b-PEO / DOTAP solution is obtained. Store it in the refrigerator for later use;

[0042] (2) Dilute the porcine epidemic diarrhea virus S protein with a buffer solution to a concentration of 50 µg / ml, and then adjust the pH value to 9.0 with an appropriate amount of sodium hydroxide and / or hydrochloric acid to obtain a porcine epidemic diarrhea virus S protein solution; , the buffer is prepared from the following components:

[0043] Sodium dihydrogen phosphate 12g

[0044] Sucrose 80g

[0045] Appropriate amount of sodium hydroxide

[0046] Hydrochloric acid amount

[0047] Glycine 0.7507g

[0048] Sodium chloride 0.2922g

[0049] Add water to 1000ml

[0050] (3) Take 250ml of the porcine epidemic diarrhea virus S protein solution prepared in step (2), add 10ml of the spare PBD-b-PEO / DOTAP solution prepared in step 1) at a rate of 10µl / min, a...

Embodiment 2

[0052] (1) Dissolve 20mg of PBD-b-PEO and 1mg of DOTAP in 10ml of tetrahydrofuran solution, and after the DOTAP lipid is dissolved, a PBD-b-PEO / DOTAP solution is obtained, which is stored in the refrigerator for later use;

[0053] (2) Dilute the porcine epidemic diarrhea virus S protein with a buffer solution to a concentration of 5 µg / ml, and then adjust the pH value to 6.0 with an appropriate amount of sodium hydroxide and / or hydrochloric acid to obtain a porcine epidemic diarrhea virus S protein solution; , the buffer solution is the same as in Example 1;

[0054] (3) Take 500ml of the porcine epidemic diarrhea virus S protein solution prepared in step (2), add 10ml of the standby PBD-b-PEO / DOTAP solution prepared in step 1) at a rate of 8µl / min, and add it at 600 rpm Stir at a certain stirring speed and mix evenly; then place the obtained solution in a homogenizer for homogenization treatment, the homogenization treatment is carried out in two stages, the first stage main...

Embodiment 3

[0056] (1) Dissolve 15mg of PBD-b-PEO and 1mg of DOTAP in 10ml of tetrahydrofuran solution. After the DOTAP lipid is dissolved, a PBD-b-PEO / DOTAP solution is obtained. Store it in the refrigerator for later use;

[0057] (2) Dilute the porcine epidemic diarrhea virus S protein with a buffer solution to a concentration of 100 μg / ml, and then adjust the pH value to 7.0 with an appropriate amount of sodium hydroxide and / or hydrochloric acid to obtain a porcine epidemic diarrhea virus S protein solution; , the buffer solution is the same as in Example 1;

[0058] (3) Take 350ml of porcine epidemic diarrhea virus S protein solution prepared in step (2), add 10ml of spare PBD-b-PEO / DOTAP solution prepared in step 1) at a rate of 12µl / min, Stir at a certain stirring speed and mix evenly; then place the obtained solution in a homogenizer for homogenization treatment, the homogenization treatment is carried out in two stages, the first stage maintains a pressure of 50±5Bar, and the sec...

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Abstract

The invention belongs to the field of animal vaccines, and particularly relates to a porcine epidemic diarrhea virus vaccine and a preparation method thereof. The vaccine consists of PBD-b-PEO / DOTAP and porcine epidemic diarrhea virus S protein; the mass ratio of the PBD-b-PEO / DOTAP to the porcine epidemic diarrhea virus S protein is (100:1)-(200:1). The porcine epidemic diarrhea virus vaccine provided by the invention can effectively deliver antigens to antigen-presenting cells, and induces antibody production and cell-mediated response; after pregnant sows are fed with the porcine epidemic diarrhea virus vaccine of the invention, the level of specific SIgA in sow milk can be significantly increased, and maintains high for a long time, and then, the specific SIgA can be passively transferred to newborn piglets through the milk, so that produced piglets can produce antibodies after weaning, and a higher serum antibody level is kept for a long time after weaning.

Description

technical field [0001] The invention belongs to the field of animal vaccines, in particular to a porcine epidemic diarrhea virus vaccine and a preparation method thereof. Background technique [0002] Porcine epidemic diarrhea (PED) is an acute, highly contagious intestinal infectious disease of porcine caused by porcine epidemic diarrhea virus (PEDV). PEDV can infect pigs of all ages, causing clinical symptoms such as vomiting, watery diarrhea, and dehydration. Piglets are the most serious infection, with a case fatality rate of 80%-100%, causing huge economic losses to the pig industry. [0003] Most of the vaccines currently on the market to prevent porcine epidemic diarrhea are traditional intramuscular injection vaccines. Due to the immature immune system of newborn piglets, the injection of vaccines cannot produce protection in time, resulting in repeated immunization failures. Passive milk-derived immunity to newborn piglets through immunization of maternal antibodie...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/215A61P31/14A61K9/127
CPCA61K9/127A61K39/12A61K2039/552A61K2039/575A61P31/14C12N2770/20034
Inventor 阮承迈高辉
Owner 天津中逸安健生物科技有限公司
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