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Method for purifying foot-and-mouth disease virus antigen by utilizing ion exchange chromatography

A technology of ion exchange chromatography and foot-and-mouth disease virus, which is applied in the field of purification of foot-and-mouth disease virus antigen by ion exchange chromatography, can solve the problems of death, side effects of immunized animals, and low purity of vaccine antigens

Pending Publication Date: 2022-03-01
天康生物制药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The vaccine antigen produced by this method has low purity and high impurity content, and the immunized animals have serious side effects or even death

Method used

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  • Method for purifying foot-and-mouth disease virus antigen by utilizing ion exchange chromatography
  • Method for purifying foot-and-mouth disease virus antigen by utilizing ion exchange chromatography

Examples

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Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1 utilizes the method for ion-exchange chromatography to purify foot-and-mouth disease AKT-III virus antigen

[0039] In this embodiment, the method for purifying FMD AKT-III virus antigen by ion-exchange chromatography comprises successively live virus suspension culture, harvesting the virus and removing cell fragments, inactivating the harvested FMD virus liquid, centrifuging and clarifying, and concentrating by ultrafiltration , depth filtration, PEG precipitation, reconstitution, ion exchange chromatography and other steps. details as follows:

[0040] BHK21 cells were cultured in suspension to a certain cell density, and then inserted into the foot-and-mouth disease AKT-III virus seed virus. After the cells were lysed, they were centrifuged to remove cell debris; the supernatant was collected and inactivated, and after the antigen was inactivated, it was clarified by deep filtration and PEG precipitation was carried out. The precipitate was redissolved...

Embodiment 2

[0052] Embodiment 2 utilizes the method for ion-exchange chromatography to purify foot-and-mouth disease O / Mya98 virus antigen

[0053] In this embodiment, the method for purifying FMD O / Mya98 virus antigen by ion exchange chromatography includes successively live virus suspension culture, collecting the virus and removing cell fragments, inactivating the harvested FMD virus liquid, centrifuging for clarification, and ultrafiltration for concentration , depth filtration, PEG precipitation, reconstitution, ion exchange chromatography and other steps. details as follows:

[0054] BHK21 cells were cultured in suspension to a certain cell density, and then inserted into the foot-and-mouth disease O / Mya98 virus seed virus. After the cells were lysed, the cell fragments were removed by centrifugation; the supernatant was collected and inactivated, and after the antigen was inactivated, it was clarified by deep filtration and PEG precipitation was carried out. The precipitate was re...

Embodiment 3

[0066] Embodiment 3 utilizes the method for ion-exchange chromatography to purify foot-and-mouth disease OHM02 virus antigen

[0067] In this embodiment, the method for purifying FMD OHM02 virus antigen by ion-exchange chromatography includes successively live virus suspension culture, harvesting the virus and removing cell fragments, inactivating the harvested FMD virus liquid, centrifuging and clarifying, ultrafiltration and concentration, deep layer Filtration, PEG precipitation, reconstitution, ion exchange chromatography and other steps. details as follows:

[0068] BHK21 cells were cultured in suspension to a certain cell density, and then inserted into the foot-and-mouth disease OHM02 virus seed virus. After the cells were lysed, the cell fragments were removed by centrifugation; the supernatant was collected and inactivated, and after the antigen was inactivated, it was clarified by deep filtration and PEG precipitation was used for precipitation. Reconstitute the rec...

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Abstract

The invention provides a method for purifying a foot-and-mouth disease virus antigen by utilizing ion-exchange chromatography, which comprises the following steps: inactivating and filtering a harvested foot-and-mouth disease virus solution, and sequentially carrying out PEG (Polyethylene Glycol) precipitation, redissolution, ion-exchange chromatography and the like. Wherein the ion exchange chromatography comprises the following steps: adopting a flow-through process, enabling a redissolved solution to pass through a chromatographic column, and collecting a flow-through component. By adopting the process, the yield of three subtype inactivated antigens of foot-and-mouth disease AKT-III, O / Mya 98 and OHM02 is more than 90%, the purity is more than 70%, the content of impure protein in the antigens is greatly reduced, more than 50% of total protein can be removed, the process is stable in amplification and short in time consumption, the process can be used for preparing high-quality foot-and-mouth disease vaccines, and the safety and effectiveness of the foot-and-mouth disease vaccines are improved.

Description

technical field [0001] The invention relates to the technical field of medicine and biology, in particular to a method for purifying foot-and-mouth disease virus antigen by ion exchange chromatography. Background technique [0002] Foot-and-Mouth Disease (FMD) is an infectious disease of cloven-hoofed animals caused by Foot-and-Mouth Disease Virus (FMDV). It is highly contagious and spreads quickly. Once it breaks out, it will cause huge economic losses. It is listed as the first of Class A zoonoses by the World Organization for Animal Health (OIE). [0003] Foot-and-mouth disease virus belongs to the Picornaviridae family and the genus Foot-and-mouth disease virus. Currently, there are 7 known serotypes, A, O, C, SAT1, SAT2, SAT3, and ASIA1, and each serotype includes many subtypes. Genetic variation among different strains can lead to antigenic differences. The main serotypes prevalent in my country are A, O and ASIA1. [0004] Vaccination is an effective way to prevent...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/09C07K1/30C07K1/18C07K1/34C07K1/36
CPCC07K14/005C12N2770/32151
Inventor 贺笋郑侃黎明蔡清波
Owner 天康生物制药有限公司
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