Asia I synthetic peptide vaccine of foot and mouth disease
A technology for synthesizing peptide vaccines and foot-and-mouth disease, which is applied in the field of vaccines and their preparation, and can solve the problems of limited gene expression products, complicated processing and disappearance of genetically engineered vaccines, etc.
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Embodiment 1
[0034] Example 1: Solid phase synthesis of synthetic peptide vaccine polypeptide antigen
[0035] The polypeptide antigen of the present invention can be prepared by ABI 433A full-automatic polypeptide synthesizer by Merrifield solid-phase synthesis method, wherein Fmoc modified amino acids are 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.
[0036] 1. Preparation of synthetic raw materials
[0037] The polypeptide antigen sequence of the synthetic peptide vaccine is shown in SEQ ID NO:1. In addition, the following polypeptides are also included:
[0038] The amino acid sequence is a peptide having an amino acid sequence in which all leucine and valine in the sequence of SEQ ID NO: 1 are substituted with norleucine and norvaline.
[0039] The amino acid sequence is a dimer of the polypeptide with the a...
Embodiment 2
[0052] Embodiment 2: the preparation of synthetic peptide vaccine
[0053] Dilute the three polypeptide antigen solutions prepared in Example 1 to 50 μg / ml with water for injection as the water phase; sterilize the 50V white oil adjuvant at 121° C. for 30 minutes, and use it as the oil phase for later use. Under the condition of 22°C, according to the volume ratio of antigen water phase and sterilized 50V as 1:1, first add the oil phase into the emulsification tank, stir at a slow speed of 100 rpm for 2 minutes, slowly add the water phase, after the addition is complete Stir for 30 minutes, then stir at a high speed of 9000 rpm for 20 minutes, then let it stand for 5 minutes, and then obtain the Asian type 1 foot-and-mouth disease virus synthetic peptide vaccine after packaging, that is, the peptide vaccine of SEQ ID NO: 1, and the peptide substituted by SEQ ID NO: 1 Vaccines and Polypeptide Vaccines of SEQ ID NO: 1.
Embodiment 3
[0054] Example 3: Study on the immune effect of synthetic peptide vaccine on guinea pigs
[0055] Healthy guinea pigs weighing 300-500 g were randomly divided into five groups, six guinea pigs in each group, and intramuscularly injected into the hind legs. See Table 1 for the injection reagents and doses of each group. The second immunization was carried out four weeks after the first immunization, and the third week was challenged after the second immunization.
[0056] Table 1: Immunization antigens and immunization doses of each test group
[0057]
[0058] 1. Detection of FMDV specific antibody
[0059] Determination of OD of serum antibody by indirect ELISA method 492 On a 96-well microtiter plate, the Asia 1 type FMDV Jiangsu virus was used as the coating antigen (1:10 dilution), the tested guinea pig serum was used as the primary antibody, and the peroxidase-labeled rabbit anti-guinea pig IgG was used as the secondary antibody. The OD value of each well was measur...
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