Vaccine composition for resisting H7 subtype avian influenza virus as well as preparation method and application of vaccine composition
A bird flu virus and vaccine composition technology, applied in the field of biopharmaceuticals, can solve problems such as poor stability and lack of immune effect of commercial whole virus vaccines
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Embodiment 1H7
[0034] Expression of embodiment 1H7 subtype avian influenza virus-like particles
[0035] 1. Carrier modification
[0036] Using the commercially available vector pFastBac I as a template, XhoI, SphI, NheI, MluI, EcoRI and XbaI restriction enzyme sites were sequentially inserted at positions 4413-4414. Add 1 μl of Dpn I enzyme to the PCR product to digest the template plasmid, take 5 μl and transform it into DH5α according to the conventional method, and the successfully transformed plasmid is named pFastBac mut.
[0037] 2. Genetic modification
[0038] The amino acid sequence shown in Seq ID No.1 of the H7N9 avian influenza virus HA gene was modified through artificial deletion, insertion and mutation: LWF 526-528 , G 531 、AIVMG 539-543 missing, I 532-533 、CVV 537-538 、MWACQ 547-548 Insertion, I536L, V545G mutations. The amino acid sequence of the modified H7N9 avian influenza virus HA gene is shown in Seq ID No.2.
[0039] 3. Construction of three expression casset...
Embodiment 2H7
[0050] The preparation of embodiment 2H7 subtype avian influenza virus-like particle vaccine
[0051] The virus-like particles harvested in Example 1 were respectively added to the mineral oil adjuvant to prepare various vaccine compositions, and the specific proportions are shown in Table 1.
[0052] Table 1 Proportion of H7 subtype avian influenza virus-like particle vaccine composition
[0053] components Vaccine 1 Vaccine 2 Vaccine 3 rBac-MNH-1 (HA content) 6log2 8log2 - rBac-MNH-2 (HA content) - - 8log2 Adjuvant (V / V%) 66% 66% 66%
Embodiment 3H7
[0054] Embodiment 3H7 subtype avian influenza virus-like particle vaccine immunogenicity test
[0055] Get 50 SPF chickens at the age of 21 days and divide them into 5 groups, 10 in each group. The first group to the third group are respectively injected with the vaccines 1 to 3 prepared in Example 2 subcutaneously in the neck, and the fourth group is injected subcutaneously with commercial products. Chemical inactivated vaccine (H5 (Re-11+Re-12), H7 (H7-Re2) trivalent inactivated vaccine, H7HA content is 8log2), immunization dose is 0.3ml, the 5th group injects 0.3ml normal saline subcutaneously, as a blank control. All experimental chickens were kept in isolation, blood was collected on the 14th and 21st day after immunization, serum was separated, and the titer of HI antibody was determined*. The test results of different HI antibodies after immunization are shown in Table 2.
[0056] Table 2 Results of immunogenicity test of H7 subtype avian influenza virus-like particle...
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Abstract
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