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

Inactive Publication Date: 2014-06-04
SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] For the detection of CSFV antibody, the currently recognized method is to use neutralization test to detect, but this method is complicated to operate, time-consuming, and requires high-quality CSF monoclonal antibody to detect, and it is difficult for ordinary laboratories to meet the above requirements

Method used

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  • Swine fever branch peptide and application thereof
  • Swine fever branch peptide and application thereof
  • Swine fever branch peptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention discloses swine fever branch peptide which connects a helper T cell epitope with a swine fever virus E2 protein B cell epitope by a branch skeleton structure. The invention further discloses application of the swine fever branch peptide as swine fever vaccine and detection antigen as detecting swine fever virus antibody. The swine fever branch peptide, as the swine fever vaccine, not only avoids immune response of an organism generating needle on a carrier, but also improves immunogenicity of synthetic peptide vaccine. Ingredients of the swine fever branch peptide are pure, have no toxic and side effect, and have no worry on biosafety, and have industrialization advantages. The swine fever branch peptide, as the detection antigen, not only is simple in preparation, but also can quickly detect the swine fever virus antibody, and has good application prospect.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a swine fever branched peptide and its application. Background technique [0002] CSFV E2 protein can induce the production of neutralizing antibodies and is the main protective antigen of CSFV (Konig et al., J Virol, 1995, 69:479-86). Some progress has been made in the identification of E2 protein epitopes. Among them, amino acids 829-837 of E2 protein are a major antigenic epitope, and it is a conserved classical classical classical epitope (Lin et al., J Virol, 2000, 74: 11619-25). The fusion product of this epitope and GST expressed in Escherichia coli induced a protective immune response (Liu et al., J Virol Meth, 2006, 134:125-9), and after tandem 4 copies, the expressed fusion protein could resist pig Pestivirus challenge (Liu et al., Vaccine, 2006, 24:7175-7180). After coupling the artificially synthesized peptide chain of the epitope with BSA, immunization of p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00A61K39/187A61P31/14G01N33/68
Inventor 童光志李国新周艳君于海
Owner SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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