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Protective agent for foot-and-mouth disease inactivated virus and preparation method of microcapsule vaccine

A technology of foot-and-mouth disease virus and foot-and-mouth disease, which is applied in the direction of antiviral agents, microcapsules, and capsule delivery, can solve problems such as lack of methods, and achieve the effects of simple formula, improved stability, and guaranteed stability

Active Publication Date: 2019-05-31
内蒙古必威安泰生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The present invention aims at the technical deficiencies of the prior art, and provides a protective agent for foot-and-mouth disease inactivated virus and a preparation method of microcapsule vaccines, so as to solve the technical problem of lacking related methods in the prior art

Method used

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  • Protective agent for foot-and-mouth disease inactivated virus and preparation method of microcapsule vaccine
  • Protective agent for foot-and-mouth disease inactivated virus and preparation method of microcapsule vaccine
  • Protective agent for foot-and-mouth disease inactivated virus and preparation method of microcapsule vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Preparation of foot-and-mouth disease microencapsulated vaccine of the present invention

[0038] 1 Preparation of inactivated antigen

[0039] 1.1 Preparation of virus liquid

[0040] 1.1.1 Preparation of type A foot-and-mouth disease virus solution

[0041] 10000L bioreactor full suspension culture of BHK-21 cells, the cell density reaches 3-5×10 6 Inoculate the cell-adapted strain of foot-and-mouth disease virus (bovine foot-and-mouth disease virus A / AKT-Ⅲ strain, prepared by Inner Mongolia Biwei Antai Biotechnology Co., Ltd.) according to the virus multiplicity of infection MOI 0.01-0.1, prepare the virus stock solution, and the stirring speed does not exceed 40rpm , culturing for 8-12h to harvest virus fluid.

[0042] 1.1.2 Preparation of O-type foot-and-mouth disease virus solution

[0043] 10000L bioreactor full suspension culture of BHK-21 cells, the cell density reaches 3-5×10 6 1 / ml, inoculate a foot-and-mouth disease virus cell-adapted strain (...

Embodiment 2

[0068] Embodiment 2 Immune effect monitoring of foot-and-mouth disease microcapsule vaccine of the present invention

[0069]The oil adjuvant vaccine prepared in Example 3.1 and the foot-and-mouth disease microcapsule vaccine prepared in Example 2.1 were immunized with a single dose of 30-45 kg, respectively, and 8 pigs were negative for FMDV antibody. 4 weeks, 8 weeks, 10 weeks, 12 weeks, 14 weeks, 18 weeks, and 20 weeks after immunization, respectively. FMDV antibody titers were determined by ELISA.

[0070] As a result, single-dose (conventional 146s content of 2-6 μg / mL) immunized pigs with foot-and-mouth disease microencapsulated vaccine, and 4-fold dose (ie, 4 times the single dose) orally immunized pigs had FMDV antibody efficacy after 8 weeks. The titer of the FMDV antibody can reach 1:1024, and the oil adjuvant vaccine can not achieve the FMDV antibody titer until 12 weeks and 16 weeks. Therefore, the foot-and-mouth disease microcapsule vaccine of the present inventi...

Embodiment 3

[0083] Embodiment 3 Embedding rate and release effect monitoring of foot-and-mouth disease microcapsules

[0084] 3.1 Determination of the encapsulation rate of foot-and-mouth disease microcapsule vaccine

[0085] Accurately weigh 0.01 g of the foot-and-mouth microcapsule vaccine prepared in Example 1, dissolve it in sterilized water filtered by a Φ0.22 μm membrane, centrifuge at a low speed of 800 rpm for 10 min, discard the supernatant, and wash away the uncoated antigen; the remaining product Precise volume into a 25mL volumetric flask. The protein content was determined by the standard curve method with the BCA protein kit (refer to the kit instructions for specific operation steps), and the encapsulation rate of the microcapsules was calculated. Calculated as follows:

[0086] Microcapsule entrapment rate (%) = (total protein content - unencapsulated protein content) / total protein content × 100%

[0087] see standard curve figure 1 , after calculation, the embedding e...

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Abstract

The invention provides a protective agent for a foot-and-mouth disease inactivated virus antigen. The protective agent consists of the following components: trehalose, raffinose, dextran, sorbitol, mannitol, inositol, xylitol, polyethylene glycol 3350, TPGS and propolis. The protective agent can prolong the preservation time of the antigen, protect the 146s antigen of the foot-and-mouth disease virus to be stable during production and cold chain transportation and is conducive to establishment of a foot-and-mouth disease virus antigen library and improvement of the low temperature storage stability of the 146s antigen in a foot-and-mouth disease virus antigen solution. The invention also provides a foot-and-mouth disease inactivated virus microcapsule vaccine containing the protective agent and the foot-and-mouth disease inactivated virus antigen as a core material and a preparation method thereof. The vaccine is orally taken by or injected to an animal for immunization, can produce ahigh titer FMDV antibody in 8 weeks and lasts longer than conventional oil adjuvant inactivated vaccines to achieve immune protection faster. The vaccine is directly injected for immunization withoutan adjuvant and can be inoculated to the animal by water drinking or feeding for prevention, and the clinical operation is convenient.

Description

technical field [0001] The invention relates to the technical field of veterinary biological products, in particular to a method for preparing a protective agent for foot-and-mouth disease inactivated virus and a microcapsule vaccine. Background technique [0002] Foot-and-mouth disease (FMD) is a severe contact infectious disease of cloven-hoofed animals caused by foot-and-mouth disease virus (FMDV), which mainly harms cattle, sheep, pigs, camels, etc., and has a very high incidence , spread extremely fast, and have great harm to the animal husbandry of various countries. Foot-and-mouth disease virus belongs to the Picornaviridae family, the genus Foot-and-mouth disease virus. Currently, there are 7 known serotypes, A, O, C, SAT1, SAT2, SAT3, and Asia1, and each serotype has many subtypes. Three serotypes are prevalent in my country, and these three serotypes are also the most pathogenic and widely distributed, namely A type, O type and Asia1 type. [0003] At present, t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/50A61K47/26A61K47/10A61K47/22A61K47/46A61K47/36A61K39/135A61P31/14
Inventor 刘月张涛何召庆张晓慧李如珩马红艳刘金萍武俊兰王永伟
Owner 内蒙古必威安泰生物科技有限公司
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