PRV (Porcine pseudorabies virus) purification method

A technology of porcine pseudorabies virus and purification method, which is applied in the fields of antiviral agents, recovery/purification, and pharmaceutical formulations, and can solve the problems of high side reaction rate, poor uniformity, and poor immune effect of porcine pseudorabies live vaccines, and achieve immune effects Good, uniform, low cost effect

Active Publication Date: 2014-07-23
TIANJIN RINGPU BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The present invention aims to solve technical problems such as high side reaction rate, poor uniformity and poor immune effect of the existing porcine pseudorabies live vaccine, and provides a porcine pseudorabies virus purification method realized by using a microfiltration clarification and purification system and an ultrafiltration concentration purification system

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] 1Operation process

[0058] 1.1 Pre-processing

[0059] 1.1.1 Assembly of the system

[0060] Under sterile conditions, the clarification and purification system and the concentration system are assembled according to the assembly requirements. A sterile 0.65μm, intact hollow fiber microfiltration membrane can be installed in the clarification system, and a sterile hollow fiber ultrafiltration membrane with a pore size of 300KD can be installed in the concentration system.

[0061] 1.1.2 System Integrity Detection

[0062] The pressure hold method checks the integrity of the purification system.

[0063] 1.1.3 System processing

[0064] Cleaning and Sterilization:

[0065] Fill the feed tank of the purification system with 0.5M NaOH solution, soak for 20 minutes, turn on the circulation pump at 300 rpm, and clean and sterilize the system for 30 minutes.

[0066] Washing and flux detection:

[0067] After the sterilization, drain the NaOH solution in the system. ...

Embodiment 2

[0110] 1) Add sterile Tween 20 into the porcine pseudorabies virus (PRV) virus solution at a ratio of 10% (v / v), and shake it;

[0111] 2) Inject the virus liquid after the oscillation treatment in step 1 into the feeding tank of the microfiltration clarification and purification system, turn on the circulation pump, and after a certain period of circulation, filter through a 0.45 μm hollow fiber microfiltration column, and collect the permeate for use;

[0112] 3) Add sterile 0.1M PBS buffer solution into the retentate in the feeding tank at a ratio of 1:1 (v / v), resuspend, turn on the circulation pump to circulate for a certain period of time, collect the permeate, and obtain the first Wash the filtrate and save it at 4-10°C for later use;

[0113] 4) Add sterile 0.1M PBS buffer solution into the retentate in the feeding tank at a ratio of 1:1 (v / v), resuspend, turn on the circulation pump to circulate for a certain period of time, collect the permeate, and obtain the second ...

Embodiment 3

[0121] 1) Add sterile Tween 20 into the porcine pseudorabies virus (PRV) virus solution at a ratio of 0.5% (v / v), and shake at 800rpm for 10min;

[0122] 2) Inject the virus liquid after shaking treatment in step 1 into the feeding tank of the microfiltration clarification and purification system, turn on the circulating pump to circulate at 400rpm for 30min, and then microfiltration through a 0.65μm hollow fiber microfiltration column to collect the permeate stand-by;

[0123] 3) Add sterile 0.1M PBS buffer solution to the retentate in the feeding tank at a ratio of 1:1 (v / v), resuspend, turn on the circulation pump and circulate at 400rpm for 30min, collect the permeate, The first washing filtrate was obtained, and stored at 4-10°C for later use;

[0124] 4) Add sterile 0.1M PBS buffer solution to the retentate in the feeding tank at a ratio of 1:1 (v / v), resuspend, turn on the circulation pump and circulate at 400rpm for 30min, collect the permeate, Obtain the second wash...

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Abstract

The invention provides a PRV (porcine pseudorabies virus) purification method. The method integrates a series of processes such as a micro-filtration clarification and purification method, an ultra-filtration concentration and purification method, and a repeated washing and filtering method, so as to increase the PRV recovery efficiency to the greatest extent and reduce the purification cost. A finished PRV concentrated solution prepared by the method is especially suitable for preparing a vaccine; compared with a porcine pseudorabies virus vaccine of the prior art, the vaccine prepared by the finished porcine pseudorabies virus concentrated solution disclosed by the invention has the advantages of high safety, good uniformity, good immune effect and the like; meanwhile, the purification method disclosed by the invention is simple and convenient in process and is relatively low in cost, and has outstanding large-scale application prospect.

Description

technical field [0001] The invention relates to a method for purifying virus, in particular to a method for purifying porcine pseudorabies virus. Background technique [0002] Pseudorabies (Porcine Pseudorabies) is an acute infectious disease caused by Pseudorabies Virus (PRV) and occurs in a variety of domestic animals and wild animals. The main symptoms are fever, itching, and encephalomyelitis. Adult pigs often show recessive infection, and pregnant sows can cause abortion, stillbirth and symptoms of respiratory diseases after infection, without itching. In addition to neurological symptoms, neonatal piglets can also damage the digestive system after infection. The disease is distributed worldwide. Pigs are not only the host of the primary infection of the pathogen, but also the long-term storage and detoxification of the virus, which plays an important role in the spread of pseudorabies. The harm caused by pseudorabies to the pig breeding industry is second only to sw...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/02A61K39/245A61P31/22
Inventor 吴全忠米娜李倬丁旭娜吕茂杰杨保收梁武李守军
Owner TIANJIN RINGPU BIO TECH
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