Synthetic peptide vaccine for treating porcine reproductive and respiratory syndrome and preparation method thereof
A technology for respiratory syndrome and synthetic peptide vaccines, which is applied in the field of peptides for synthetic peptide vaccines, can solve the problems of short immune protection period, multiple immunizations, and strong virulence, and achieve high levels of immune response, high safety, and efficacy Good results
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Embodiment 1
[0035] Example 1 : Solid Phase Synthesis of Polypeptide Antigens for Synthetic Peptide Vaccines
[0036] The polypeptide antigen of the present invention can be prepared by ABI 433A automatic polypeptide synthesizer by Merrifield solid-phase synthesis method, wherein 9-fluorenylmethoxycarbonyl (Fmoc) modified amino acid is used, and the solid phase carrier is Rink Amide MBHA resin. The production process usually includes solid-phase synthesis of polypeptide antigens, cleavage of polypeptides, purification of antigens and sterilized storage.
[0037] (1) Preparation of synthetic raw materials
[0038] Polypeptide antigens for polypeptide vaccines are synthesized, and their amino acid sequences are shown in SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3 or SEQ ID NO.4.
[0039] Prepare appropriate Fmoc-modified amino acids according to the above peptide antigen sequence and 1 mmol synthesis scale, and add them to the corresponding Cartridge (vial containing amino acids). Also weigh th...
Embodiment 2
[0050] Example 2 : Preparation of Synthetic Peptide Vaccines
[0051] The polypeptide solution prepared in Example 1 was diluted to 200 μg / ml with water for injection, and used as the water phase; the SEPPIC MONTANIDE ISA 50V oil adjuvant was sterilized at 121°C for 30 minutes, and used as the oil phase for later use. At 22°C, according to the volume ratio of antigen water phase and sterilized 50V oil adjuvant of 1:1, first add the oil phase into the emulsification tank, stir at a slow speed of 100 rpm for 2 minutes, then slowly add the water phase, Stir for 30 minutes after the addition, then stir at a high speed of 9000 rpm for 20 minutes, then let stand for 5 minutes, and then obtain the synthetic peptide vaccine against porcine reproductive and respiratory syndrome virus after subpackaging, namely SEQID NO.1 polypeptide vaccine, SEQ ID NO. ID NO.2 polypeptide vaccine, SEQ ID NO.3 polypeptide vaccine, SEQID 4 polypeptide vaccine.
Embodiment 3
[0052] Example 3 : Efficacy Test of Synthetic Peptide Vaccines
[0053] (1) Vaccines and experimental animals
[0054] Four batches of PRRS synthetic peptide vaccine plus inactivated vaccine (purchased from China Animal Husbandry Co., Ltd. Chengdu Biopharmaceutical Factory) and adjuvant (50V oil) were set up as six groups in total, in which the inactivated vaccine was used as a positive control, the adjuvant was used as a negative control, and the experimental animals were 60 healthy pigs at the age of 40 days were tested negative for PRRSV antigen and antibody. See Table 1 for specific grouping.
[0055] Table 1 Vaccine grouping situation
[0056] group
Number of test animals (head)
Immunization dose (ml / head)
Group A
10
2
Group B
SEQ ID NO.1 polypeptide vaccine
10
2
Group C
SEQ ID NO.2 polypeptide vaccine
10
2
Group D
SEQ ID NO.3 polypeptide vacci...
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