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Preparation method of porcine reproductive and respiratory syndrome vaccines by utilizing bioreactor

A bioreactor, respiratory syndrome technology, applied in the field of preparation of porcine reproductive and respiratory syndrome vaccines, can solve the problems of unstable product quality, large differences between product batches, limiting vaccine quality and output, etc., to ensure production process parameters and product quality, reduce batch-to-batch variation, and improve the effect of toxigenic titer

Active Publication Date: 2014-07-09
RINGPU (BAODING) BIOLOGICAL PHARMACEUTICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Cell and virus culture conditions are difficult to control during production, resulting in unstable cell and vaccine quality, large batch-to-batch differences, low automation, and high labor intensity, which greatly limits the quality and yield of vaccines
[0004] With the rapid development of biotechnology, the traditional production methods have been difficult to meet the needs of the market, and the use of bioreactors is a good choice
However, there are still many problems in the production of vaccines using reactors: for example, due to the different states of inoculated cells and unscientific inoculation methods, there are large differences between vaccine batches and unstable product quality; due to unscientific nutrient supply methods, cell nutrition is lacking and growth is not good. Uniformity; due to the difficulty in the expansion of the cell seed chain, it is difficult to realize the expansion and cultivation of cells; and due to the cumbersome operation, increased pollution probability, and energy consumption caused by cleaning and sterilization, many problems are difficult to enumerate

Method used

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  • Preparation method of porcine reproductive and respiratory syndrome vaccines by utilizing bioreactor
  • Preparation method of porcine reproductive and respiratory syndrome vaccines by utilizing bioreactor
  • Preparation method of porcine reproductive and respiratory syndrome vaccines by utilizing bioreactor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Production of porcine reproductive and respiratory syndrome live vaccine (R98 strain) by AP20C type torrent perfusion bioreactor

[0047]In the AP20C type torrent perfusion bioreactor used in this example, the perfusion bag 6) has a volume of 5 L, the torrent bag 5) has a volume of 15 L, and the perfusion bag (6) contains 150 g of polyester fiber paper carrier 8); theoretically effective cultivation Volume (harvesting liquid volume) is 10L.

[0048] figure 1 It is a schematic diagram of the composition of the liquid circulation system of the torrent perfusion bioreactor; in which, the cells and viruses used for seedling production are all grown in the perfusion bag, and the nutrient solution (cell growth solution, cell maintenance solution) passes through the external peristaltic pump (the first peristaltic pump 1. The second peristaltic pump 2) circulates in the perfusion bag and the torrent bag.

[0049] The seed virus is the R98 strain of porcine reprodu...

Embodiment 2

[0072] Example 2 Producing Porcine Reproductive and Respiratory Syndrome Live Vaccine (R98 Strain) with AP200 Type Torrent Perfusion Bioreactor

[0073] In the AP200 type torrent perfusion bioreactor used in this example, the perfusion bag 6 has a volume of 50 L, the torrent bag 5 has a volume of 150 L, and the perfusion bag 6 contains 2000 g of polyester fiber paper carrier 8; the theoretical effective culture volume (harvesting liquid Volume) is 100L.

[0074] A. Preparation:

[0075] Two torrent perfusion bioreactors, AP20C and AP200, were selected to prepare for cell culture.

[0076] B. Production work, the specific steps are as follows:

[0077] a, the cultivation of cells for seedling preparation:

[0078] According to the method of Example 1, the Marc-145 cells cultured in the AP20C bioreactor, on the 4th day, when the daily glucose consumption reached 3.8g / d / L, the cells in the perfusion bag 6) were digested with 0.025% trypsin, Prepare 17L of cell suspension, con...

Embodiment 3

[0095] Example 3 Comparative Experiment of Three Methods of Harvesting Virus Liquid

[0096] 1. Materials and methods

[0097] Materials such as equipment, seed poison are with embodiment 2;

[0098] The experiment was divided into three groups, all cultivating cells for making seedlings, inoculating virus, and amplifying porcine reproductive and respiratory syndrome virus according to the same method in Example 2, but the three groups used different methods to harvest the virus liquid, and the harvested virus liquid was all The volume and titer of the virus fluid were measured according to conventional methods.

[0099] The first group adopts one-time harvesting method: see embodiment 2;

[0100] The second group adopts the multiple harvesting method: the first harvest at 24 hours, the harvest once every 6 hours thereafter, and the last harvest at 48 hours, a total of 5 harvests; the first 4 harvests, each harvest of 70-90% of the total volume of virus liquid %, and su...

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Abstract

A kind of method utilizing bioreactor to prepare porcine reproductive and respiratory syndrome vaccine, it utilizes a torrent perfusion bioreactor as cultivating tool, and preparation is carried out according to following steps: a. the cultivation of the cell that makes seedling; b. virus inoculation and Cultivation; c. virus liquid harvest; d. vaccine preparation and other processes. The invention has the advantages of simple operation, low pollution probability, significantly reduced production cost, large vaccine output, uniform and stable quality, and excellent immune effect. The whole production process does not involve other biological safety and public health problems, and is suitable for large-scale production.

Description

technical field [0001] The invention relates to a method for preparing a porcine reproductive and respiratory syndrome vaccine by using a bioreactor, in particular to a method for producing a porcine reproductive and respiratory syndrome vaccine by using a torrent perfusion bioreactor, which belongs to the field of biotechnology. Background technique [0002] Porcine reproductive and respiratory syndrome (PRRS) is an infectious disease caused by porcine reproductive and respiratory syndrome virus (PRRSV), which seriously endangers the pig industry. At present, the main way to control PRRS is to use vaccines for immune prevention. Therefore, high-titer virus liquid is a very important link in vaccine production. [0003] The production of porcine reproductive and respiratory syndrome vaccine is mainly through the cultivation of cells in spinner bottles, and then the propagation of porcine reproductive and respiratory syndrome virus. It is difficult to control the cell and vi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/12A61P31/14C12M3/00
Inventor 邱贞娜郑朝朝杨保收郁宏伟刘涛
Owner RINGPU (BAODING) BIOLOGICAL PHARMACEUTICAL CO LTD
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