A kind of vaccine composition and its preparation method and application
A vaccine composition and antigen technology, which can be used in pharmaceutical formulations, medical preparations containing active ingredients, virus antigen components, etc., and can solve problems such as simultaneous immune interference
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Embodiment 1
[0031] The preparation of embodiment 1 swine fever virus antigen liquid
[0032] 1. The highly sensitive ST cells (purchased from ATCC) containing 0.125% trypsin and 0.03 %EDTA digestion solution, digest and disperse, inoculate cell culture flasks at a suitable density after counting cells, add 1.5-5% FBS MEM cell culture solution, and add seed poison at the same time according to M.O.I. The inoculation dose is M.O.I.=0.1-0.6, more preferably the inoculation dose is M.O.I.=0.2-0.4, and cultured in an incubator at 34-37°C, more preferably at a temperature of 34-35°C.
[0033] 2. After three days of cultivation, the first time of poisoning is carried out. After the poisoning, the cell maintenance solution of 1.5-3% FBS is added. After that, the poison is collected every 2 days, and the poison can be collected 5 times continuously. After harvesting the virus, the antigens were mixed and stored at -20°C.
[0034] 3. Mix and sample the virus fluid harvested five times before perf...
Embodiment 2
[0035] The preparation method of embodiment 2 highly pathogenic mutant strain antigen of PRRS
[0036] 1. Passage and culture of cells for seedling production: Disperse and passage Marc145 or MA104 cell lines with EDTA-trypsin digestion solution, and continue to culture with cell growth medium.
[0037] 2. The propagation of cytotoxic species: the virus liquid of PRRS HuN4-F112 strain (commercially available) is inserted into the cell bottle that has grown into a good cell monolayer by the dose of M.O.I.=0.001-0.01, and the dose of poisoning is further optimized M.O.I. = 0.005-0.01. After 1 hour of adsorption, add cell maintenance solution to the cell line monolayer and continue to culture. Harvest when 70-80% of the cells have lesions. Save as below, take a small amount for inspection of semi-finished products, the virus content of the antigen solution is 10 6.5 TCID 50 / ml, in accordance with the national standard inspection are in compliance with the regulations.
[0038...
Embodiment 3
[0040] Example 3 Double vaccine composition prepared after different ratios of swine fever, porcine highly pathogenic PRRS (HuN4-F112, JXA1-R, TJM-F92) antigens and swine fever virus antigens
[0041] The hog fever virus antigen prepared by embodiment 1 (virus content 10 6.5 TCID 50 / ml), three kinds of porcine PRRS virus antigens prepared in embodiment 2 carry out proportioning according to the antigen content of table 1 respectively, then mix with the freeze-drying protection agent of equal amount, make porcine PRRS, swine fever double See Table 1 for the live vaccine vaccine composition.
[0042] Table 1 Different ratios and groups of PRRS antigens and CSF antigens
[0043]
[0044] Mix the above antigens in sequence, then add a mixture of 2wt% gelatin and 15wt% lactose in a volume ratio of 1:1 as a freeze-drying protective agent, stir and mix for 1 hour, carry out aseptic subpackaging, and store at 2-8°C ,spare.
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