Canine parvovirus proliferation method

A technology of canine parvovirus and cells, which is applied in the fields of virus culture and vaccine antigen preparation, can solve the problems of low culture efficiency and achieve the effects of stable product quality, easy realization, and improved proliferation efficiency

Inactive Publication Date: 2015-11-11
TIANJIN RINGPU BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention aims at the technical defects of the prior art, and provides a canine parvovirus propagation method to solve the technical problem of low culture efficiency existing in the prior art

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] 1. Passage and culture of production cells

[0026] Take well-grown F81 cells, pass through EDTA-trypsin cell dispersion solution (EDTA-trypsin cell dispersion solution is PBS solution containing 0.25% by mass volume fraction of specification 1:250 trypsin, 0.02% EDTA; wherein specification 1:250 Trypsin (trypsin containing 250 activity units per gram of trypsin) was digested and passaged, and the pH was adjusted to 7.4 with 100 units / ml each of 90% DMEM solution, 10% bovine serum, penicillin sodium and streptomycin. The cell growth medium was continuously cultured at 37°C to form a good monolayer of F81 cells, which was used for continuous passage or inoculated in a bioreactor for microcarrier suspension culture after passage.

[0027] 2. Propagation of seed poisons for production

[0028] After the F81 cells have been cultured for 2 days to form a single layer, pour out the cell growth medium, digest with the EDTA-trypsin cell dispersion solution, mix the dispersion ...

Embodiment 2

[0036] A method for multiplying canine parvovirus. In the method, F81 cells are used as host cells to propagate canine parvovirus, and a torrent bioreactor is used as a cell culture device.

[0037] Specifically, the F81 cells are first cultivated in a torrent bioreactor, and then the canine parvovirus is inoculated into the cell culture medium to proliferate the parvovirus.

[0038] The culture of the F81 cells is to cultivate the F81 cells in a rapid flow bioreactor by means of microcarrier suspension culture.

[0039] The method specifically includes the following steps:

[0040] 1) Add cell culture medium to the rapid flow bioreactor, with 0.1×10 6 F81 cell culture was inoculated at the ratio of cells / mL culture solution, and there were microcarriers in the reactor;

[0041] 2) When the cell density reaches 1×10 6 cells / mL culture medium, continue to culture the cells for 4 hours, then insert canine parvovirus into the culture medium at an inoculum of 0.01 MOI, and then...

Embodiment 3

[0049] A method for multiplying canine parvovirus. In the method, F81 cells are used as host cells to propagate canine parvovirus, and a torrent bioreactor is used as a cell culture device.

[0050] Specifically, the F81 cells are first cultivated in a torrent bioreactor, and then the canine parvovirus is inoculated into the cell culture medium to proliferate the parvovirus.

[0051] The culture of the F81 cells is to cultivate the F81 cells in a rapid flow bioreactor by means of microcarrier suspension culture.

[0052] The method specifically includes the following steps:

[0053] 1) Add cell culture medium to the rapid flow bioreactor at 1×10 6 F81 cell culture was inoculated at the ratio of cells / mL culture solution, and there were microcarriers in the reactor;

[0054] 2) When the cell density reaches 10×10 6 cells / mL of culture medium, continue to culture the cells for 6 hours, then inoculate the culture medium with canine parvovirus at an inoculum of 0.03 MOI, and th...

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PUM

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Abstract

The invention provides a canine parvovirus proliferation method. According to the method, F81 cells are creatively taken as host cells of canine parvovirus, so that virus proliferation is facilitated especially; a riptide-type bioreactor is adopted, so that compared with conventional spinner bottle modes, processes are easier to control, and products are more stable in quality; a microcarrier culture mode is adopted, so that a culture solution can be utilized fully, growing characteristics of adherent cells are maintained, and high-density culture effect is realized. In addition, aiming at own characteristics of the F81 cells and the canine parvovirus and culture environment of the bioreactor, specific culture conditions are matched, and components of the culture solution, inoculation amount and condition parameters are designed in an optimized manner, so that canine parvovirus proliferation efficiency is improved remarkably. Protruding technological effect is realized through an ingenious technological concept, and the method is low in cost and easy in realization and has outstanding popularization prospect.

Description

technical field [0001] The invention relates to the technical field of virus cultivation, and further relates to the technical field of vaccine antigen preparation, in particular to a method for multiplying canine parvovirus. Background technique [0002] Canine parvovirus (CPV) belongs to the parvovirus genus of the family Parvoviridae, and mainly infects dogs, especially puppies. After healthy dogs are infected with CPV, the virus mainly attacks intestinal epithelial cells and cardiomyocytes, showing gastrointestinal symptoms and myocarditis symptoms respectively. Canine parvovirus is highly contagious, and the mortality rate of dogs after infection is high. For canine parvovirus infection cases, clinically, symptomatic treatment is mainly combined with hyperimmune serum and monoclonal antibody treatment, which is not ideal in terms of curative effect and cost. In general, vaccination is the fundamental measure to deal with the virus. This involves the proliferation techn...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C12R1/93
Inventor 孙晨苏建东杨保收付旭彬梁武
Owner TIANJIN RINGPU BIO TECH
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