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Porcine reproductive and respiratory syndrome virus (PPRSV) purification method

A technology of breathing disorder and purification method, which is applied in the field of virus purification, can solve the problems of poor uniformity of virus vaccines and poor immune effect, and achieve the effect of highlighting the prospect of large-scale application, low cost and high purity

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

[0006] The present invention aims to solve the technical problems such as poor uniformity and poor immune effect of existing porcine reproductive and respiratory syndrome virus vaccines, and provides a porcine reproductive and respiratory syndrome virus vaccine that is realized by using a microfiltration clarification and purification system and an ultrafiltration concentration and purification system. Virus purification method

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] 1Operation process

[0057] 1.1 Pre-processing

[0058] 1.1.1 Assembly of the system

[0059] 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.

[0060] 1.1.2 System Integrity Detection

[0061] The pressure hold method checks the integrity of the purification system.

[0062] 1.1.3 System processing

[0063] Cleaning and Sterilization:

[0064] Fill the feed tank of the 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.

[0065] Washing and flux detection:

[0066] After the sterilization, drain the NaOH solution in the system. Fill the feed...

Embodiment 2

[0133] 1) Add sterile Tween 20 to the porcine reproductive and respiratory syndrome virus (PPRSV) virus solution at a ratio of 10% (v / v), and shake it;

[0134] 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 circulation pump, and after a certain period of circulation, pass through a 0.45 μm hollow fiber microfiltration column to microfiltration, and collect the permeate for use;

[0135] 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 for use;

[0136] 4) Add sterile 0.1M PBS buffer solution to the retentate in the feed tank that has been processed in step 3) at a ratio of 1:1 (v / v), resuspend, turn on the circulation pump for a certain period of time, and collect Permeat...

Embodiment 3

[0144] 1) Add sterile Tween 20 into the porcine reproductive and respiratory syndrome virus (PPRSV) virus solution at a ratio of 0.5% (v / v), and shake at 1000rpm for 10min;

[0145] 2) Inject the virus liquid after the shaking treatment in step 1 into the feeding tank of the microfiltration clarification and purification system, turn on the circulating pump to circulate at 500rpm for 30min, and then microfiltration through a 0.65μm hollow fiber microfiltration column to collect the permeate stand-by;

[0146] 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 500rpm for 30min, collect the permeate, Obtain the first wash filtrate stand-by;

[0147] 4) Add sterile 0.1M PBS buffer solution to the retentate in the feed tank that has been processed in step 3) at a ratio of 1:1 (v / v), resuspend, turn on the circulation pump for a certain period of time, and collect Permeate ...

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Abstract

The invention provides a porcine reproductive and respiratory syndrome virus (PRRSV) purification method. The method comprehensively adopts a series of processes, such as microfiltration clarification purification process, ultrafiltration concentration purification process, repeated filter wash process and the like, maximally enhances the recovery efficiency of PRRSV, and lowers the purification cost. The PPRSV concentrated solution finished product prepared by the method is especially suitable for preparing a vaccine; and compared with the PPRSV vaccine in the prior art, the vaccine prepared from the PPRSV concentrated solution finished product has the advantages of high safety, high purity, favorable uniformity and favorable immunization effect. The method has the advantages of simple technique and lower cost, and has outstanding popularization prospects.

Description

technical field [0001] The invention relates to a method for purifying virus, in particular to a method for purifying porcine reproductive and respiratory syndrome virus. Background technique [0002] Porcine reproductive and respiratory syndrome, also known as porcine blue-ear disease, is a viral infectious disease that endangers the pig industry. In the summer and autumn of 2006, pig "high fever" epidemics occurred in some provinces in southern my country. The Ministry of Agriculture actively organized scientific research personnel to jointly tackle the problem, and identified the pig PRRS variant virus as the main pathogen, which was named "highly pathogenic pig PRRS". Until today, the epidemic situation is still severe. Like other viral diseases, there is currently no effective therapeutic drug for the treatment of PRRS. Immunization is the most effective way to prevent and control the disease. Currently commercialized PRRS Vaccines are mainly attenuated live vaccines p...

Claims

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Application Information

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IPC IPC(8): C12N7/02A61K39/12A61P31/14C12R1/93
Inventor 米娜吴全忠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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