High-pathogenicity porcine reproductive and respiratory syndrome heat-resistant protective agent live vaccine and preparation method thereof

A technology of heat-resistant protective agent and respiratory syndrome, applied in biochemical equipment and methods, vaccines, antiviral agents, etc., can solve problems such as insufficient production automation, large measurement deviation, and pollution caused by hand manipulation, and solve energy problems. Inconvenient waste and storage, high aging resistance, and less potency loss

Active Publication Date: 2020-08-28
哈尔滨元亨生物药业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At the same time, in the preparation method, the measurement deviation is relatively large through manual operation, and the hand pinch during the manual preparation process will also cause pollution, the efficiency is low, and it is impossible to quickly carry out mass production. In order to optimize the problems existing in the above preparation method, by improving the preparation method method, and optimize the autoclave used in the vaccine preparation process to solve the problems of insufficient production automation, easy contamination of raw materials and low production efficiency in the existing preparation methods. The present invention proposes a highly pathogenic pig Reproductive and respiratory syndrome heat-resistant protective agent live vaccine and preparation method thereof

Method used

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  • High-pathogenicity porcine reproductive and respiratory syndrome heat-resistant protective agent live vaccine and preparation method thereof
  • High-pathogenicity porcine reproductive and respiratory syndrome heat-resistant protective agent live vaccine and preparation method thereof
  • High-pathogenicity porcine reproductive and respiratory syndrome heat-resistant protective agent live vaccine and preparation method thereof

Examples

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Effect test

preparation example Construction

[0042] A preparation method of a highly pathogenic porcine reproductive and respiratory syndrome heat-resistant protective agent live vaccine, the preparation method comprising the following steps:

[0043] S1: Take gelatin, tryptone, sucrose, polyvinylpyrrolidone K30, and mannitol, and add them to the corresponding leaky tanks 2 in the autoclave, and control the switch to make the above ingredients in different leaky tanks 2 press Sequentially leak into the reaction tank 3 below, and all dissolve in the water for injection in the reaction tank 3, and replenish the water for injection to the required amount after fully dissolving;

[0044] S2: Start the stirring mechanism, fully mix and stir the mixture of the components in the reaction tank 3, adjust the environment in the reaction tank 3 of the autoclave, control the temperature at 120°C-125°C, and the pressure at 14-15 pounds, and carry out Autoclaving requires the sterilization time to be maintained for 20-30 minutes, then...

Embodiment approach

[0053] As an embodiment of the present invention, the upper surface of the slider 11 is fixedly connected with a second motor 32, and the output shafts of the second motor 32 pass through the slider 11 and extend to the lower position of the slider 11; The output shafts of the two motors 32 are all fixedly connected with the third telescopic rod 33; A connecting column is fixedly connected; the surface of the connecting column is provided with evenly arranged air holes 35; when working, by setting the second motor 32, the rotation of the second motor 32 will drive the rotation of the third telescopic rod 33, and the third telescopic rod 33 will be rotated through the third telescopic rod. The expansion and contraction of rod 33 can be disconnected automatically with connecting column, when the 3rd telescopic rod 33 is connected between connecting column, the 3rd telescopic rod 33 can drive agitating plate 34 to rotate, and the rotation of agitating plate 34 can promote reaction...

Embodiment 1

[0062] The vaccine prepared by formula 1 heat-resistant protective agent in Table 1 is diluted to 1 portion / ml with serum-free DMEM cell culture medium, and then 10 times of serial dilution is made, and 10 -3 、10 -4 、10 -5 、10 -6 A total of 4 dilutions were inoculated into 96-well Marc-145 cell culture plates that had grown into a good monolayer and the cell culture medium was discarded. Each dilution was inoculated into 6 wells, 0.1ml per well, and a normal cell control was set at the same time. Set at 37°C, containing 5% CO 2 After adsorption in the incubator for 1 hour, add 0.1ml of DMEM cell culture medium containing 4% newborn bovine serum to each well, and place at 37°C, containing 5% CO 2 Cultivate and observe in the incubator for 5 days, and calculate TCID according to the Reed-Muench method 50 , the virus content per serving should be ≥10 5.5 TCID 50 , to measure the change of virus content before and after freeze-drying, and to test the shelf life of the vaccin...

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Abstract

The invention belongs to the technical field of biology, and particularly relates to a high-pathogenicity porcine reproductive and respiratory syndrome heat-resistant protective agent live vaccine anda preparation method thereof. The heat-resistant protective agent live vaccine is high in aging resistance, the loss rate after storage for 7 days at 37 DEG C does not exceed 1 titer, and after the vaccine is stored for 18 months at 2-8 DEG C, the titer of the vaccine is reduced a little, so that the problems of energy waste and storage inconvenience caused by the fact that the vaccine needs to be subjected to frozen storage in the transportation and storage processes are effectively solved; through a high-pressure sterilization cabinet used in the preparation method, by arranging a shell, aleakage tank and a reaction tank, automatic weighing of raw materials of a heat-resistant protective agent is effectively achieved, rapid dissolving and uniform mixing of all the raw materials are achieved, meanwhile, the interior of the reaction tank can be automatically heated and pressurized, and finally the heat-resistant protective agent is prepared; and the preparation process is comprehensively and mechanically carried out, the cleanliness is relatively high, the influence of human factors is reduced, and the efficiency is relatively high.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a highly pathogenic porcine reproductive and respiratory syndrome heat-resistant protective agent live vaccine and a preparation method thereof. Background technique [0002] In the prior art, highly pathogenic porcine reproductive and respiratory syndrome attenuated vaccines that are common on the market need to be stored below -15°C, which will increase the cost of transportation and storage of the vaccine. The heat-resistant protective agent of the present invention is live The vaccine has good anti-aging effect. After being stored at 37°C for 7 days, the titer loss is less than 1 titer. It can be stored at 2-8°C for 18 months, which will greatly reduce transportation costs and energy waste. [0003] At the same time, in the preparation method, the measurement deviation is relatively large through manual operation, and the hand pinch during the manual preparation process will also...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/12A61K47/42A61P31/14A61L2/07
CPCA61K39/12A61K47/42A61K2039/552A61L2/0023A61L2202/21A61P31/14C12N2770/10034
Inventor 叶阳任德强张健麻昌姣宋新刚张立恒孙博郭伟孙雪亭尹辉
Owner 哈尔滨元亨生物药业有限公司
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