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Solution for large-scale production of foot-and-mouth disease virus-like particle antigens, and purification and assembly method

A technology of foot-and-mouth disease virus and particle antigen, which is applied in the field of biomedicine, can solve the problems of low production efficiency, unsuitability for large-scale production, and high production cost, and achieve the effects of short time consumption, short purification time, and large processing capacity

Inactive Publication Date: 2020-04-10
SICHUAN HUAPAI BIO PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this method is not suitable for large-scale production due to high production cost and low production efficiency.

Method used

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  • Solution for large-scale production of foot-and-mouth disease virus-like particle antigens, and purification and assembly method
  • Solution for large-scale production of foot-and-mouth disease virus-like particle antigens, and purification and assembly method
  • Solution for large-scale production of foot-and-mouth disease virus-like particle antigens, and purification and assembly method

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Experimental program
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Embodiment 1

[0043] The preferred embodiment of the present invention provides a purification and assembly method for large-scale production of foot-and-mouth disease virus-like particle antigens. The specific steps are as follows:

[0044] (1) Preparation of solutions: prepare Buffer A, Buffer B, Buffer C and Buffer D solutions respectively;

[0045] Buffer A solution: containing 18mM sodium dihydrogen phosphate, 5mM disodium hydrogen phosphate, 300mM sodium chloride, 2% Trition-X100.

[0046] Buffer B solution: containing 18mM sodium dihydrogen phosphate, 5mmM disodium hydrogen phosphate, 300mM sodium chloride, 80mM imidazole.

[0047] Buffer C solution: containing 18mM sodium dihydrogen phosphate, 5mmM disodium hydrogen phosphate, 300mM sodium chloride, 400mM imidazole.

[0048] Buffer D solution: containing 30mM Tris-HCl, 300mM sodium chloride, 2mM calcium chloride, 6% glycerin.

[0049] (2) Treatment of fermented liquid: the fermented liquid of foot-and-mouth disease virus recombina...

Embodiment 2

[0059] The preferred embodiment of the present invention provides a purification and assembly method for large-scale production of foot-and-mouth disease virus-like particle antigens. The specific steps are as follows:

[0060] (1) Preparation of solutions: prepare Buffer A, Buffer B, Buffer C and Buffer D solutions respectively;

[0061] Buffer A solution: containing 15mM sodium dihydrogen phosphate, 2mM disodium hydrogen phosphate, 100mM sodium chloride, 1% Trition-X100.

[0062] Buffer B solution: containing 15mM sodium dihydrogen phosphate, 2mmM disodium hydrogen phosphate, 100mM sodium chloride, 50mM imidazole.

[0063] Buffer C solution: containing 15mM sodium dihydrogen phosphate, 2mmM disodium hydrogen phosphate, 100mM sodium chloride, 300mM imidazole.

[0064] Buffer D solution: containing 10mM Tris-HCl, 200mM sodium chloride, 1mM calcium chloride, 3% glycerin.

[0065] (2) Treatment of fermented liquid: the recombinant protein fermented liquid of foot-and-mouth dis...

Embodiment 3

[0075] The preferred embodiment of the present invention provides a purification and assembly method for large-scale production of foot-and-mouth disease virus-like particle antigens. The specific steps are as follows:

[0076] (1) Preparation of solutions: prepare Buffer A, Buffer B, Buffer C and Buffer D solutions respectively;

[0077] Buffer A solution: containing 20mM sodium dihydrogen phosphate, 10mM disodium hydrogen phosphate, 500mM sodium chloride, 5% Trition-X100.

[0078] Buffer B solution: containing 20M sodium dihydrogen phosphate, 10mmM disodium hydrogen phosphate, 500mM sodium chloride, 100mM imidazole.

[0079] Buffer C solution: containing 20mM sodium dihydrogen phosphate, 10mmM disodium hydrogen phosphate, 500mM sodium chloride, 500mM imidazole.

[0080] Buffer D solution: containing 50mM Tris-HCl, 500mM sodium chloride, 5mM calcium chloride, 10% glycerin.

[0081] (2) Treatment of fermented broth: The fermented broth of foot-and-mouth disease virus recombi...

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Abstract

The invention discloses a solution for large-scale production of foot-and-mouth disease virus-like particle antigens and a purification and assembly method, belonging to the technical field of biological medicines. With the method, foot-and-mouth disease virus-like particles with high purity and high assembly rate is purified and prepared from foot-and-mouth disease virus recombinant protein fermentation liquor expressed by escherichia coli. The method is simple in technological process, convenient to operate and high in production efficiency; the purification yield of the foot-and-mouth disease virus-like particles reaches 90% or above, and the in-vitro assembly rate of the particles reaches 67%; and the method is particularly suitable for industrial production of foot-and-mouth disease virus-like particle antigens expressed by escherichia coli.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a solution, purification and assembly method for large-scale production of foot-and-mouth disease virus-like particle antigens. Background technique [0002] Foot-and-mouth disease (FMD) is an acute, highly contagious animal disease that can spread rapidly over long distances and is caused by the Foot-and-Mouth Disease Virus (FMDV), which is mainly caused by cloven-hoofed animals. It is a severe infectious disease that poses the greatest threat to the development of animal husbandry. The World Organization for Animal Health (OIE) lists it as an infectious disease that must be reported. At present, whole-virus inactivated vaccines are mainly used to prevent and control FMD at home and abroad, but the effect of epidemic prevention is not satisfactory. Due to the high production cost, biosafety problems of loose virus caused by incomplete inactivation during the vac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/09C07K1/36C07K1/34C07K1/22C12P21/06
CPCC07K14/005C12N2770/32123C12N2770/32151C12P21/06
Inventor 李峰于作邱文英李妍谢建勇吴越龚文波方鹏飞严成何承勇庄万敏朱遥王俊朱旗严欢沈皓月李捷余谦
Owner SICHUAN HUAPAI BIO PHARMA
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