Method for preparing porcine circovirus through basket reactor fermentation

Abstract

The invention discloses a method for preparing porcine circovirus through basket reactor fermentation. The method for preparing the porcine circovirus through the basket reactor fermentation is characterized by comprising the following steps: (1), preparing the host cell species; (2), preparing a basket reactor before inoculation; (3), performing host cell propagating culture; and (4), performingvirus proliferating culture: emptying cell growing liquid in the reactor, supplementing cell maintaining liquid with the volume equal to that of the emptied cell growing liquid, inoculating the porcine circovirus, 36-48h after inoculation, harvesting a virus solution for the first time, after harvesting the virus solution, adding the cell maintaining liquid with the volume equal to that of the harvested virus solution into the reactor, propagating, reproducing, and harvesting the virus again, circularly repeating for multiple times until the sugar consumption of cells in the virus solution tobe harvested is detected to be reduced down to 0.5g / L / day, and harvesting the last porcine circovirus solution in the batch. The method is simple and feasible; and by the method, the production cost can be greatly reduced, the unit titer of a porcine circovirus antigen is significantly improved, and the harvest of the virus solution antigen in each fermentation batch is increased.

Description

technical field [0001] The invention relates to a production method for large-scale amplification of porcine circovirus, in particular to a method for preparing porcine circovirus by fermenting in a basket reactor. Background technique [0002] Porcine circovirus (PCV) can be divided into two types according to serotype: porcine circovirus type I (abbreviated as PCV1) and porcine circovirus type II (abbreviated as PCV2). PCV1 is not pathogenic but can produce Serum antibodies, PCV2 can have a strong susceptibility to pigs, and it is the cause of weaned piglet multisystem wasting syndrome (PMWS), porcine respiratory disease syndrome (PRDC), porcine skin and nephrotic syndrome (PDNS), etc. The main pathogen of the disease, PCV has 7 open reading frames (ORFs), among which ORF2 encodes the Cap protein of the virus, which constitutes the nucleocapsid of the virus. There is a common antigenic determinant on the Cap protein of PCV1 and PCV2, but there is no antigenic cross-reacti...

AI Technical Summary

Problems solved by technology

Although compared with the traditional spinner bottle culture process, it has the advantages of less land occupation, less energy consumption, less labor, easier expansion of production scale, higher virus toxicity, and stable product quality, but its production process has the following problems: ① There are problems of contact probability and compatibility between cells and microcarriers, and it is not easy to inoculate cells uniformly; ② The cell growth medium needs to be emptied during virus inoculation, and microcarriers will be lost when the medium is changed; Impurities such as cell debris in the microcarrier are cleaned, so they cannot be recycled and can only be used once, and the microcarrier is expensive, resulting in high production costs; ④ The growth state of the cells on the microcarrier is an extended form, close to a planar structure, and the virus is inoculated Finally, the cell death process is synchronous, that is, most of the cell death time is close, and in order to improve the efficiency of virus infection, an incubator whose main component is glucosamine needs to be added to the cell culture medium to promote the virus's infection of cells , so the virus can only be harvested once

Although the NBS basket bioreactor has been used in cell culture, however, different cells and viruses have different requirements on the culture environment and culture process conditions. Only when the conditions are suitable, the number of cultured cells or viruses Many, good activity, and high virus toxicity