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Emulsion breaking method for aftosa oil emulsion inactivated vaccine

A technology of inactivated vaccines and oil emulsions, applied in the field of quality inspection of foot-and-mouth disease vaccines, can solve the problems of high capital expenditure, high requirements for experimental facilities, and long time consumption, and achieve high recovery rate, good antigen separation effect, and stability.

Inactive Publication Date: 2012-03-21
JINYUBAOLING BIO PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The existing vaccine quality standards stipulate that the efficacy test must use this animal for testing. Since the country implements a 100% enhanced immunization policy, it is difficult to select susceptible animals for testing, and animal challenge has high requirements for experimental facilities (BSL3 level laboratory), time-consuming (more than one month), and large capital expenditure
If the serum neutralization test is used to select susceptible animals, it is technically difficult to exclude non-susceptible animals with cellular immunity, and irregular test data often occurs in practice, which affects the accuracy of the test

Method used

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  • Emulsion breaking method for aftosa oil emulsion inactivated vaccine
  • Emulsion breaking method for aftosa oil emulsion inactivated vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] The demulsification method of embodiment one foot-and-mouth disease oil emulsion inactivated vaccine

[0016] Take 100ml of commercial foot-and-mouth disease vaccine emulsified with ISA 206 adjuvant, put it into a 250ml separating funnel, add 6-20ml of demulsifier n-amyl alcohol (or choose any one of benzyl alcohol, chloroform, and n-hexanol), and fully suspend for 1 Minutes or more; put the separatory funnel into a refrigerator at 4°C, and let it stand for more than 30 minutes, oil and water can be seen to separate; separate about 34ml of the water phase layer, which contains the 146S antigen.

Embodiment 2

[0017] The recovery and content determination test of the 146S antigen after embodiment two foot-and-mouth disease oil emulsion inactivated vaccine demulsification

[0018] Get 4 parts of foot-and-mouth disease oil emulsion inactivated vaccine samples, respectively numbered as 1#-4#, according to the method in embodiment 1 for demulsification; get the water phase obtained after demulsification, that is, 146S antigen solution, according to the invention patent of application number 200910092779 The described sucrose density gradient ultraviolet light quantitative method detects the content of 146S antigen, and the specific steps are as follows:

[0019] 1. Concentration of 146S antigen solution: Ultracentrifuge containing 146S antigen solution, resuspend to 1 / 10 of the original volume with PBS solution to obtain concentrated antigen solution, and put it in a refrigerator at 2-8°C for later use;

[0020] 2. Prepare 15%-45% sucrose gradient: prepare 5 tubes of 15%-45% uniform lin...

Embodiment 3

[0035] 146S antigen content determination in embodiment three foot-and-mouth disease oil emulsion inactivated vaccines

[0036] The content determination method of 146S antigen in the foot-and-mouth disease oil emulsion inactivated vaccine is as follows:

[0037] (1) get the commercial foot-and-mouth disease inactivated vaccine of 100ml ISA 206 adjuvant emulsification, demulsify by the method in embodiment one, obtain containing 146S antigen aqueous phase about 34ml;

[0038] (2) Detect the 146S antigen content in the aqueous phase according to the sucrose density gradient ultraviolet light quantitative method described in Example 2 (X 0 ) was 0.78 μg / ml, and the corrected value (X) of 146S antigen content was further obtained as 0.58 μg / ml;

[0039] (3) According to the formula Y=-1.0343X 2 +2.244X-0.0082, the calculated 146S antigen content (Y) in the vaccine is 0.94 μg / ml.

[0040]It is known that the amount of 146S antigen added to the vaccine sample during preparation ...

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Abstract

The invention relates to an emulsion breaking method for an aftosa oil emulsion inactivated vaccine used in an aftosa vaccine quality detection technology. The method comprises the following steps of: fully and uniformly mixing an emulsion breaking agent with an aftosa oil emulsion inactivated vaccine; standing; separating oil from water; and distributing aftosa virus particles into a water phase, wherein the emulsion breaking agent is selected from any of benzyl alcohol, n-amyl alcohol, chloroform and hexyl alcohol serving as organic solvents. Due to the adoption of the method, the completeness of aftosa virus particles 146S is not broken basically, emulsion is broken rapidly and simply, the antigen 146S recovery rate is high, and the stability is high; and the method is particularly suitable to be taken as a method for monitoring the quality of commercial aftosa vaccines for aftosa vaccine producing enterprises and the national quality test departments.

Description

technical field [0001] The invention relates to a key step in the quality detection technology of the foot-and-mouth disease vaccine, in particular to a demulsification method of the foot-and-mouth disease oil emulsion inactivated vaccine. Background technique [0002] Foot-and-mouth disease (FMD) is an acute, severe, and contagious infectious disease caused by foot-and-mouth disease virus (FMDV) in cloven-hoofed animals, mainly harming pigs, cattle, sheep, etc. 70 Breeding domestic animals and wild animals are also susceptible to human beings, which are extremely harmful to animal husbandry. Foot-and-mouth disease virus is an RNA virus, composed of 4 structural proteins (VP1, VP2, VP3 and VP4) and 7 non-structural proteins in an icosahedron. The centrifugal sedimentation coefficient of intact virus particles is 146S. In general, it is believed that vaccines prepared with whole virus particles (ie, the 146S antigen) work best. The production process of the commonly used i...

Claims

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Application Information

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IPC IPC(8): B01D17/04B01D17/025
Inventor 韩润林刘国英刘玉梅徐师军范秀丽
Owner JINYUBAOLING BIO PHARMA CO LTD
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