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Preparation method of foot-and-mouth disease vaccines

A foot-and-mouth disease vaccine and antigen technology, which is applied in the preparation of foot-and-mouth disease vaccine and the purification process of foot-and-mouth disease vaccine cell culture liquid, can solve the problems of increasing the risk of cross-contamination, large antigen loss, and increasing impurity load.

Inactive Publication Date: 2015-08-12
北京必威安泰科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As product molecular titers increase, higher cell clumps and larger quantities of product present challenges for downstream purification steps, higher cell clumps cause difficulties during clarification and sterile filtration, and higher product concentrations generally result in increased Impurity load
At present, the inactivated foot-and-mouth disease vaccine only uses centrifugation technology to remove limited cell fragments. The foot-and-mouth disease virus cell culture treated by this method contains a large amount of non-target protein residues. , the centrifugal cleaning procedure slows down the plant's ability to produce different strains of FMD virus and greatly increases the risk of cross-contamination between production runs

Method used

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  • Preparation method of foot-and-mouth disease vaccines
  • Preparation method of foot-and-mouth disease vaccines

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] ——Preparation of foot-and-mouth disease type O and Asia1 bivalent inactivated vaccines

[0042] Will BHK 21 Suspension cells were respectively inoculated with the cell culture medium of foot-and-mouth disease virus ONXC / 92 strain and Asia1 / JSL / GSZY / 06 strain, concentrated and purified by the following method, and the inactivated two virus antigen solutions were mixed and diluted, and then added 206 The adjuvant is mixed and emulsified to make a vaccine for the prevention of bovine O-type and Asia1-type foot-and-mouth disease. BHK 21 Cells were purchased from China Veterinary Drug Administration.

[0043] 1. Provide foot-and-mouth disease vaccine cell culture fluid and depth filter device.

[0044] (1) The foot-and-mouth disease virus used for vaccine production of the present invention is ONXC / 92 strain and Asia1 / JSL / GSZY / 06 strain, purchased from Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences. The two strains were respectively inocu...

Embodiment 2

[0055] ——Preparation of Foot-and-Mouth Disease Type O Bivalent Inactivated Vaccine

[0056] BHK21 suspension cells were inoculated with O / Mya98 / XJ / 2010 and O / GX / 09-7 strain cell culture fluid respectively, and the two kinds of virus antigen solutions after concentration, purification and inactivation were mixed and diluted by the following method, and then Add 206 adjuvant to mix and emulsify to make a vaccine, which is used for the prevention of porcine O-type foot-and-mouth disease.

[0057] 1. Provide foot-and-mouth disease vaccine cell culture fluid and depth filter device.

[0058] (1) The virus used in this vaccine is foot-and-mouth disease virus O / Mya98 / XJ / 2010 strain and O / GX / 09-7 strain, purchased from Lanzhou Veterinary Research Institute. The two strains were respectively inoculated with BHK21 cell suspension culture to obtain the antigen solution.

[0059] (2) The device used for deep filtration uses diatomite as a filter aid, and the feed liquid is in contact wi...

Embodiment 3

[0070] ——Quality inspection of foot-and-mouth disease ONXC / 92 strain and Asia1 / JSL / GSZY / 06 bivalent inactivated vaccine

[0071] The test is divided into two groups, the group A adopts the original purification method to prepare the vaccine, and the group B adopts the technology used in the present invention to prepare the vaccine.

[0072] 1. The mensuration of effective antigen content of vaccine: detect foot-and-mouth disease 146S antigen content with sucrose density gradient method, get 10 samples respectively before and after each group of purification, get average value, compare two experimental groups 146S antigen content, as shown in table 1, result It shows that the loss of the antigen purified by the present invention is small, and the content of the effective antigen obtained is high.

[0073] Table 1 Comparison of viral antigen content in vaccines prepared by two processes

[0074] group

146S before purification (μg / ml)

Purified 146S (μg / ml)

e...

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Abstract

The invention provides a preparation method of foot-and-mouth disease vaccines. The method comprises the following steps: (1) filtering a cell culture fluid of viruses by using a deep filter device at a flow velocity of 300-500 L / m<2> / hr; (2) carrying out ultrafiltration and concentration on antigen liquid by using a hollow fiber column or membrane envelope; (3) enabling a virus antigen obtained by using the purification and concentration method to pass through DEAE-Sepharose FF and Sephawse6FF two-step chromatographic columns; (4) carrying out inactivation by using BEI, and blocking by using sodium thiosulfate; and (5) carrying out emulsification by using a 206 adjuvant. Due to vaccines prepared by using the method, immune animals can achieve a good immune effect, less adverse reaction and high safety.

Description

technical field [0001] The invention relates to a preparation method of a foot-and-mouth disease vaccine, which is a novel purification process of a foot-and-mouth disease vaccine cell culture solution. It belongs to the field of veterinary biological product manufacturing. Background technique [0002] Foot-and-mouth disease is an acute, highly contagious, febrile infectious disease caused by foot-and-mouth disease virus (FMDV) in cloven-hoofed animals (Bachrach HL.Foot-and-mouth disease virus[J].Ann Rev Microbiol, 1968,22:201-244.) (Domingo E, Baranowski E, Escarrnrs C, et al. Foot-and-mouth disease virus [J]. Comp. Immun. Microbiol-Infect. Dis, 2002, 25: 297- 308.), it is very harmful to the breeding industry, and it is listed as the first of Class A infectious diseases by the International Veterinary Bureau, and it is one of the infectious diseases that the World Organization for Animal Health (OIE) must report. FMDV belongs to the Picornaviridae family and the genus F...

Claims

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Application Information

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IPC IPC(8): A61K39/135A61P31/14
Inventor 何召庆赵颖慧张涛李如珩田普厚赵炳武刘彦涛康斌王永伟张澍
Owner 北京必威安泰科技有限公司
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