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
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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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