A kind of vaccine composition and its preparation method and application
A technology of vaccine composition and antigen, which is applied in the field of vaccine composition, can solve the problems of triple combination vaccine of Haemophilus parasuis that does not yet exist, increase strength, and stress response of pig herds, so as to reduce costs and reduce vaccinations The number of times, the effect of enhancing the immune effect
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Embodiment 1
[0039] The preparation of embodiment 1 classical swine fever antigen, porcine circovirus antigen and Haemophilus parasuis antigen
[0040] 1 Preparation of swine fever antigen
[0041] 1.1 The ST cells (purchased from ATCC) that are highly sensitive to classical swine fever virus (the attenuated strain of classical swine fever rabinized by the China Veterinary Drug Administration, preservation number AV1412) grown into a good monolayer were treated with 0.125% trypsin and 0.03% Digest and disperse the EDTA digestion solution, inoculate the cell culture bottle after counting the cells, add 1.5%-5% FBS MEM cell culture medium, and add seed poison at the same time according to M.O.I. M.O.I.=0.1-0.6, more preferably the inoculated dose is M.O.I.=0.2-0.4, placed in a 34-37°C incubator for cultivation, further preferably the cultivation temperature is 34-35°C.
[0042] After three days of cultivation, the first time of poisoning is carried out. After the poisoning, the cell mainten...
Embodiment 2
[0059] The preparation of embodiment 2 swine fever, porcine circular ring and Haemophilus parasuis triple mixed vaccine composition
[0060] Preparation of 1 para-Hemophilus suis and porcine circular double inactivated vaccine
[0061] 1.1 Preparation of preservatives
[0062] 1% (w / v) thimerosal aqueous solution: 1g of thimerosal is dissolved in 100ml of purified water, and autoclaved at 121°C for 30 minutes for later use.
[0063] 1.2 Preparation of diluent
[0064] Sterile PBS buffer solution: Dissolve 8g sodium chloride, 0.25g potassium chloride, 3.63g disodium hydrogen phosphate, 0.24g potassium dihydrogen phosphate in 900ml purified water, then dilute to 1L, autoclave at 121°C for 30min spare.
[0065] 1.3 Vaccine adjuvant treatment
[0066] Sterilization of the Gel01 adjuvant: transfer the Gel01 adjuvant into a sterilizable container, and autoclave at 121° C. for 30 minutes for later use.
[0067] 1.4 Matching seedlings
[0068] Through aseptic operation, the conc...
Embodiment 3
[0076] Example 3 Different antigen content swine fever, porcine circular and Haemophilus parasuis triple mixed vaccine efficacy test
[0077] 1 test material
[0078] Vaccine 1 (Haemophilus parasuis 10) prepared in embodiment 2 7 CFU / capacity, swine fever virus 10 4.0 TCID 50 / touquan, porcine circovirus 10 4.0 TCID 50 / head), vaccine 2 (Haemophilus parasuis 10 8 CFU / capacity, swine fever virus 10 5.0 TCID 50 / touquan, porcine circovirus 10 5.0 TCID 50 / head), vaccine 3 (Haemophilus parasuis 10 9 CFU / capacity, swine fever virus 10 6.0 TCID 50 / touquan, porcine circovirus 10 6.0 TCID 50 / head), vaccine 4 (Haemophilus parasuis 10 10 CFU / capacity, swine fever virus 10 6.0 TCID 50 / touquan, porcine circovirus 10 6.0 TCID 50 per head) and vaccine 5 (Haemophilus parasuis 2×10 10 CFU / capacity, swine fever virus 10 6.0 TCID 50 / touquan, porcine circovirus 10 6.0 TCID 50 / toufen).
[0079] 3-4 week-old weaned piglets free of antibodies to CSF, Circus porcine,...
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