Swine fever branch peptide and application thereof
A technology of branched peptides of swine fever and swine fever virus, applied in the field of bioengineering, can solve the problems of difficult access in the laboratory, complicated operation, time-consuming, etc., and achieve the effects of easy control of production quality, improvement of immunogenicity, and simple ingredients
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Embodiment 1
[0040] The branched peptide E29 and the linear peptide E26 shown below were artificially synthesized by conventional methods in the field of chemical synthesis. The purity of the synthetic peptides is above 70%. Dilute the synthetic peptides with sterile PBS to a concentration of 1 microgram per microliter, then mix the same amount of Freund's adjuvant with the peptide solution, fully emulsify, and use it as an immunogen in immunized animals.
[0041] Branched Peptide-E29
[0042] Linear peptide-E26 ISISEIKGVIVHKIEGILFKKCTAVSPTTLRTEVVK
[0043] Select 15 healthy rabbits weighing about 2 kg, and divide them into 3 groups, 5 rabbits in each group. Rabbits No. 8, 9, and 10) were immunized with branched peptide E29, and the third group (rabbits No. 11, 12, 13, 14, and 15) were immunized with PBS as a negative control. The peptide immunogen immunization dose was 50 micrograms each time, with a total volume of 500 microliters, injected into the skin at multiple points, and immu...
Embodiment 2
[0046] Peptide immunogens were prepared as in Example 1 and used to immunize animals. Select 20 healthy piglets that were 30 days old and negative for CSF antibody, and divide them into 4 groups, 5 pigs in each group. The first group was immune to linear peptide E26, the second group was immune to branched peptide E29, and the remaining 2 groups were immune to attenuated CSF vaccine. (HCLV) and PBS, as positive and negative controls. The peptide immunogen immunization dose is 200 micrograms each time, the total volume is 1 milliliter, multi-point intramuscular injection, immunization 2 times, with an interval of 2 weeks. The first emulsification with Freund's complete adjuvant and the second emulsification with Freund's incomplete adjuvant. Before immunization and after immunization, blood was collected every week to separate serum, and the serum antibody level was detected with IDEXX swine fever antibody detection kit.
[0047] The results showed that 2 weeks after the seco...
Embodiment 3
[0049] The synthetic peptides E26 and E29 were diluted with the coating solution, and the final concentration was 0.5ug / 100ul. The two synthetic peptides were used as the detection antigens of indirect ELISA respectively, and 5 samples of CSFV positive serum, 1 CSFV negative serum and Porcine pseudorabies virus (PRV) positive serum, porcine blue ear disease virus (PRRSV) positive serum, porcine circovirus type 2 (PCV2) positive serum, porcine parvovirus (PPV) positive serum, porcine transmissible gastroenteritis virus ( TGEV) positive serum, 1 part of porcine epidemic diarrhea virus (PEDV) positive serum, detect the specificity of synthetic peptide E26 and E29 as the detection antigen of swine fever antibody.
[0050] The operating procedure of indirect ELISA experiment is as follows:
[0051] 1. Coating: Dilute the synthetic peptide (0.5ug / 100ul) with the coating solution, add to the microtiter plate, 100ul per well, and coat overnight at 4°C.
[0052] 2. Washing: wash (200u...
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