O-type foot-and-mouth disease virus-like particle antigen, vaccine composition containing O-type foot-and-mouth disease virus-like particle antigen, and preparation method and application of vaccine composition
A technology of foot-and-mouth disease virus and vaccine composition, which is applied in the field of O-type foot-and-mouth disease virus-like particle antigens, can solve the problems of no formation, poor vaccine immune effect, and inability to effectively exert the immunogenicity of VLPs vaccines, and achieve good stability and maintenance The effect of antibody titer and immune effect
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Embodiment 1
[0045] Example 1 Expression of O-type Ind strain foot-and-mouth disease virus-like particles
[0046] 1. Construction of expression vector
[0047] The VPO gene fragment shown in the sequence table SEQ ID NO.1, the VP3 gene fragment shown in the sequence table SEQ ID NO.2, and the VP1 gene fragment shown in the sequence table SEQ ID NO.3 were synthesized by Jinweizhi Company, respectively, and pBLUE -T Vector vector connection, connect the successfully connected recombinant clones to the pET28a vector after the same restriction digestion, The positive clone pET28a-VP0-VP3-VP1 inserted into O-type Ind strain foot-and-mouth disease virus VPO, VP3, VP1 genes was obtained. Transform the ligated plasmid into CaCl 2 The prepared DH5α competent cells were spread on the kanamycin-resistant solid LB medium, and when the monoclonal colony was clearly visible, the single clone was picked into the LB liquid medium containing kanamycin, 37 C, 230 rev / min, cultivate overnight for 12 hour...
Embodiment 2
[0055] The preparation of embodiment 2 O type Ind strain foot-and-mouth disease virus-like particle vaccine composition
[0056] Take the virus-like particles prepared in Example 1 and slowly add them to the adjuvant. During the process of adding, stir continuously with an emulsifier with a rotation speed of 800 rpm for 12 minutes, mix well, and store at 4° C., which is the foot-and-mouth disease virus-like particle vaccine composition. See Table 1 for specific ratios. The adjuvants suitable for the present invention may be adjuvants known to those skilled in the art. In this example, the adjuvant ISA 206 (Seppic, France) was selected.
[0057] Table 1 O-type Ind strain foot-and-mouth disease virus-like particle vaccine composition ratio
[0058] components Vaccine 1 Vaccine 2 Vaccine 3 FMD antigen (μg / ml) 160 200 240 ISA 206 Adjuvant (V / V%) 50% 50% 50%
Embodiment 3
[0059] Example 3 The immunogenicity test of the O-type Ind strain foot-and-mouth disease virus-like particle vaccine composition
[0060] Select 20 healthy susceptible piglets with a weight of about 40kg, negative for type O foot-and-mouth disease virus antigen and antibody, and divide them into 4 groups randomly, 5 pigs in each group. Groups 1-3 are vaccine 1, vaccine 2, and vaccine 3 immunization groups prepared in Example 2 of the present invention, respectively, and group 4 is a control group. The way of immunization in the immunized group was intramuscular injection of 2 ml in the neck, and the control group was immunized with the same amount of PBS. Blood was collected from each pig before vaccine immunization, and blood was collected on the 7th, 14th, 21st, and 28th days after immunization.
[0061] The collected sera were tested for related antibodies using the Type O Ind strain foot-and-mouth disease antibody ELISA detection kit. The results showed that the antibodi...
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