Purification and concentration method of Seneca virus

A virus concentration and virus technology, which is applied in the field of purification and concentration of Seneca virus, achieves the effects of mild process, convenient operation and high antigen recovery rate

Inactive Publication Date: 2020-05-19
JINYUBAOLING BIO PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the disease has formed a trend of global spread. Once a large-scale outbreak occurs, it will inevitably cause immeasurable economic losses.

Method used

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  • Purification and concentration method of Seneca virus
  • Purification and concentration method of Seneca virus
  • Purification and concentration method of Seneca virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1: Purification and concentration of type A Seneca virus liquid

[0040] (1) Sterilization and balance

[0041]Use 100L of 0.5mol / L NaOH solution to clean the hollow fiber concentration system for 30-60min, and then soak in 0.5mol / L NaOH solution overnight.

[0042] (2) Integrity detection

[0043] Clean the hollow fiber concentration system with purified water. When the pH value of the eluate is ≤7.0, apply clean compressed air to the hollow fiber concentration system to a pressure of 0.10MPa for leak detection. If there is no continuous bubble escaping within 1min, it is qualified. Use 40mmol / 100 L of PBS (pH=7.5-7.6) solution in 100 L is used to clean the hollow fiber concentration system that has passed the integrity test.

[0044] (3) Pretreatment of Seneca virus liquid

[0045] Take the Seneca virus liquid (very toxic), centrifuge at 12,000 rpm / min for 20 min in a batch high-speed centrifuge, discard the precipitate, and take the supernatant for lat...

Embodiment 2

[0051] Embodiment 2: Purification and concentration of type A Seneca inactivated venom

[0052] (1) Inactivation of Seneca virus

[0053] Add β-propiolactone at 0.25‰ of the volume of the virus solution, incubate at 4°C for 24 hours, and hydrolyze at 37°C for 2 hours to inactivate the virus.

[0054] (2) Sterilization and balance

[0055] Use 100L of 0.5mol / L NaOH solution to clean the ultrafiltration membrane package system in a circular manner for 30-60min, and then soak overnight in 0.5mol / L NaOH solution after cleaning. Use 100 L of 40 mmol / L PBS (PH=7.5-7.6) solution to clean the ultrafiltration membrane capsule system, and stop cleaning when the pH value of the eluate is ≤7.5.

[0056] (3) Pretreatment of Seneca virus liquid

[0057] Take the inactivated Seneca virus liquid (attenuated) in step (1), use a cartridge filter (0.2μm≤filter membrane pore size≤0.65μm) to filter and clarify, and take the filtrate for later use.

[0058] (4) concentrated

[0059] Pass 400L ...

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Abstract

The invention discloses a concentration method of Seneca virus. The method comprises pretreatment of a Seneca virus liquid, and concentration after the pretreatment, wherein the concentration sequentially comprises to filtration concentration, filter wash and filtration concentration. The invention relates to an SVV antigen concentration and purification method suitable for large-scale industrialproduction. The method subjects an SVV antigen to concentration and purification by combining tangential flow filtration and filter wash, and has the characteristics of convenient operation, easy amplification, mild and stable process, high antigen recovery rate and the like. The method can be used for concentration and purification of live viruses and inactivated viruses, has an antigen recoveryrate of greater than 80% and a protein removal rate of greater than 90%, and can prepare a purified SVV antigen or provide raw materials for subsequent further fine purification of the antigen.

Description

technical field [0001] The invention relates to the technical field of preparing virus vaccines, in particular to a method for purifying and concentrating Seneca virus. Background technique [0002] Seneca virus (Seneca Valley virus, SVV), also known as Seneca virus A (SVA), is a single-stranded positive-sense RNA virus without an envelope, and is a member of the Senecavirus genus of the family Picornaviridae. ) is a newly discovered virus that can cause porcine vesicular-like symptoms in recent years, and is considered to be the main cause of porcine primary vesicular disease (PIVD). After the virus infects animals (especially pigs), it can cause vesicular lesions on the snout, crown and hooves of breeding sows and boars, with occasional diarrhea symptoms, and adult pigs are usually subclinical. What is even more frightening is that the virus can cause the death of newborn piglets, and the mortality rate of newborn piglets at the age of 1-3 days can reach 30%-70%. After p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/02C12R1/93
CPCC12N7/00C12N2770/32051
Inventor 陈光达陈九连田志辉刘国英张燕红史琳凯路荣齐志涛泰鹏
Owner JINYUBAOLING BIO PHARMA CO LTD
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