Multi-component subunit vaccine for African swine fever, and preparation method and application of multi-component subunit vaccine
A subunit vaccine and African swine fever technology, applied in the field of animal vaccines and veterinary biological products, can solve the problems of complex pathogenic mechanism, biological safety hidden dangers, immune interference, and poor immune effect, so as to achieve no immune interference and prevent And protect infection, meet the effect of industrial application
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Embodiment 1
[0026] Embodiment 1: the preparation of African swine fever virus CD2V protein, P54 protein, P30 protein and P72 protein
[0027] 1.1 Preparation of African swine fever virus CD2V protein: refer to the preparation method of African swine fever CD2V protein in the invention patent of application number 201910004596.5 or 201910069838.9 or the preparation method in other patents or literatures to prepare African swine fever CD2V protein, and the full-length can be prepared according to actual needs African swine fever CD2V protein or CD2V-Fc fusion protein.
[0028] 1.2 Preparation of African swine fever virus p54 protein: refer to the published literature on the expression of African swine fever virus p54 protein and the preparation of monoclonal antibody (Feng Chunyan et al., 2016, China Animal Quarantine) or the prokaryotic expression and analysis of African swine fever virus p54 recombinant protein. Its monoclonal antibody preparation (Feng Yao, 2014, master's thesis of Yangzho...
Embodiment 2
[0031] Embodiment 2: the preparation of African swine fever subunit vaccine
[0032] The consumables and materials used for the preparation of vaccines need to undergo aseptic treatment in advance, and the preparation process is completed in a biological safety cabinet or other instruments or environments that can ensure that the entire preparation process is sterile.
[0033] 2.1 Vaccine preparation (taking the mixture of four proteins as an example)
[0034] Water phase preparation: According to the CD2V, p54, TF-P72, TF-P30 protein content in the vaccine, use PBS (or normal saline) to dilute the CD2V, p54, TF-P72, TF-P30 protein to an appropriate concentration, which is the water phase;
[0035] Preparation of oil phase: According to the total amount of prepared vaccine, according to the weight ratio of antigen phase and adjuvant is 1:1, and the volume ratio is 46:54, measure an appropriate amount of ISA 201 VG adjuvant;
[0036] Emulsification: Preheat both the water phas...
Embodiment 3
[0038] Embodiment 3: African swine fever subunit vaccine immunization experiment
[0039] 3.1 Vaccine preparation: Prepare proteins and vaccines according to the methods of Examples 1 and 2, and the specific vaccine information is shown in Table 1 below:
[0040] Table 1
[0041]
[0042] 3.2 Immunization experiment: Screen 40 piglets aged 28-35 days (CSFV, PCV2, PRRSV antigen antibody negative), randomly divide into 8 groups, 5 piglets in each group, one group serves as blank control group, and the other 7 groups are immunized with vaccine 1 and vaccine respectively 2. Vaccine 3, Vaccine 4, Vaccine 5, Vaccine 6, Vaccine 7. The blank control group was intramuscularly injected with 1ml of normal saline each time. In addition, 1ml of the corresponding vaccine was intramuscularly injected into the 7 groups of immunization groups each time, and a booster immunization was given three weeks after the initial immunization. Serum was collected before immunization, before the seco...
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