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Porcine epidemic diarrhea virus vaccine and preparation method thereof

A porcine epidemic diarrhea and virus vaccine technology, applied in the field of porcine epidemic diarrhea virus vaccine and its preparation, can solve the problems of difficult preparation of porcine epidemic diarrhea virus inactivated vaccine, decreased piglet resistance, unfavorable piglet protection, etc. Significant difference in titer, good immune protection, and the effect of increasing the level of specific SIgA

Active Publication Date: 2020-09-25
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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  • Porcine epidemic diarrhea virus vaccine and preparation method thereof
  • Porcine epidemic diarrhea virus vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0042] (2) Dilute the S protein of porcine epidemic diarrhea virus with a buffer solution to a concentration of 50 µg / ml, and then adjust the pH 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 by the following components:

[0043] Sodium dihydrogen phosphate 12g

[0044] Cane sugar 80g

[0045] Sodium hydroxide amount

[0046] Hydrochloric acid

[0047] Glycine 0.7507g

[0048] Sodium chloride 0.2922g

[0049] Add water to 1000ml

[0050] (3) Take 250 ml of the porcine epidemic diarrhea virus S protein solution prepared in step (2), and add 10 ml of the spare PBD-b-PEO / DOTAP solution prepared in step 1) at a rate of 10 µl / min, at 700 rpm Stir at the mixing speed ...

Embodiment 2

[0052] (1) Dissolve 20mg PBD-b-PEO and 1mg DOTAP in 10ml tetrahydrofuran solution. After the DOTAP lipid is dissolved, the PBD-b-PEO / DOTAP solution will be obtained. Store in the refrigerator for later use;

[0053] (2) Dilute the S protein of porcine epidemic diarrhea virus with a buffer solution to a concentration of 5 µg / ml, and then adjust the pH 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 is the same as in Example 1;

[0054] (3) Take 500 ml of the porcine epidemic diarrhea virus S protein solution prepared in step (2), and add 10 ml of the spare PBD-b-PEO / DOTAP solution prepared in step 1) at a rate of 8 µl / min. Stir at different stirring speeds and mix evenly; then place the resulting solution in a homogenizer for homogenization treatment. The homogenization treatment is carried out in two stages. The first stage maintains the pressure at 50±5Bar, and the second st...

Embodiment 3

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

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

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

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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, and specifically relates to a porcine epidemic diarrhea virus vaccine and a preparation method thereof. Background technique [0002] Porcine epidemic diarrhea (PED) is an acute and highly contagious porcine intestinal infectious disease caused by porcine epidemic diarrhea virus (PEDV) infection. PEDV can infect pigs of all ages, causing animals to have clinical symptoms such as vomiting, watery diarrhea and dehydration. Piglets have the most serious infection, with a mortality rate of 80%-100%, causing huge economic losses to the pig industry. [0003] At present, most of the vaccines on the market to prevent swine epidemic diarrhea are traditional intramuscular vaccines. Due to the imperfect development of the immune system of newborn piglets, the injection of vaccines cannot produce protection in time, resulting in repeated immunization failures. By immunizing the sow, the maternal antibodies produced...

Claims

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Application Information

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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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