CN-121991181-A - Epstein-Barr virus 18 type key epitope peptide and application thereof

Abstract

The invention relates to screening of key Epitope, in particular to an Epstein-Barr virus 18 type key Epitope peptide and application thereof, and the results of an ELISA (enzyme-linked immunosorbent assay) experiment show that Epitope 1 (SEQ ID NO: 1) can be effectively combined with E18-VP1 antiserum, and the combining capacity is obviously stronger than that of blank control and negative result Epitope 2, so that the invention screens out 1 Epitope peptide based on the whole sequence of the Epstein-Barr virus 18-Barr virus 1, and has important guiding significance for research and development of an Epstein-Barr virus 18 vaccine.

Inventors

• WU CHUNCHEN
• WU JIE
• XIA JIANBO
• JIANG SHUAI
• SUN TAO
• GUO MIN

Assignees

• 湖北省妇幼保健院（湖北省妇女儿童医院）
• 中国药科大学

Dates

Publication Date

20260508

Application Date

20251231

Claims (7)

1. 1. The critical antigen epitope peptide of the Epstein-Barr virus 18 is characterized in that the amino acid sequence of the antigen epitope peptide is shown as SEQ ID NO. 1.
2. 2. A fusion protein or polypeptide is characterized in that the fusion protein or polypeptide comprises at least one epitope peptide as set forth in claim 1, and the amino acid sequence of the epitope peptide is shown as SEQ ID NO. 1.
3. 3. The use of an epitope peptide according to claim 1 for the preparation of a vaccine for the treatment of an epstein barr virus.
4. 4. The use according to claim 3, wherein the serotype of the epstein barr virus is epstein barr virus type 18.
5. 5. The use of the epitope peptide of claim 1 for preparing an epstein-barr virus type 18 detection reagent.
6. 6. A vaccine comprising the epitope peptide of claim 1 or the fusion protein or polypeptide of claim 2, and a pharmaceutically acceptable carrier.
7. 7. The vaccine of claim 6, wherein the pharmaceutically acceptable carrier is selected from one or more of an adjuvant, an excipient.

Description

Epstein-Barr virus 18 type key epitope peptide and application thereof Technical Field The invention relates to critical epitope screening, in particular to an Epstein-Barr virus 18 type critical epitope peptide and application thereof. Background Human orphan enterocytopathic virus (Enteric Cytopathogenic Human Orphan Virus, ECHO), abbreviated as Epstein-Barr virus, belongs to enterovirus type B (EV-B) of enterovirus genus of picornaviridae, and is a single-stranded positive strand RNA virus. The Epstein-Barr virus is related to rash, hand-foot-and-mouth disease, aseptic meningitis, encephalitis, paralysis, myocarditis and other diseases, and is mainly transmitted through a fecal-oral route, so that the Epstein-Barr virus can infect people of all ages, especially infants. Global enterovirus epidemics and distribution overview shows that epstein barr virus 18 (E18) is one of the most frequently used serotypes with highest prevalence of EV-B and is an important viral pathogen in children. E18 infection can cause a variety of diseases, such as a range of diseases from mild fever to potentially fatal sterile meningitis, encephalitis, paralysis, myocarditis, etc. At present, clinical infection aiming at the Epstein-Barr virus mainly adopts symptomatic treatment, and no effective therapeutic drugs or preventive vaccines have been developed. There has been no study reporting the epitope sequence of the epstein barr virus 18 and no vaccine developed based thereon. Disclosure of Invention The first object of the present invention is to provide an amino acid sequence of an epitope peptide derived from the Ekkoviral 18-VP1 protein. The invention also aims at providing an application for predicting and screening the epitope peptide. The technical scheme is that the key epitope peptide of the Epstein-Barr virus 18 is characterized in that the amino acid sequence of the epitope peptide is shown as SEQ ID NO. 1. A fusion protein or polypeptide is characterized in that the fusion protein or polypeptide comprises at least one epitope peptide as set forth in claim 1, and the amino acid sequence of the epitope peptide is shown as SEQ ID NO. 1. The application of the antigen epitope peptide in preparing the vaccine for treating the Epstein-Barr virus. The use, wherein the serotype of the epstein barr virus is epstein barr virus type 18. The application of the antigen epitope peptide in preparing an Epstein-Barr virus 18 type detection reagent. A vaccine comprising the epitope peptide of claim 1 or the fusion protein or polypeptide of claim 2, and a pharmaceutically acceptable carrier. The vaccine is characterized in that the pharmaceutically acceptable carrier is selected from one or more of an adjuvant and an excipient. The invention has the following research ideas: 1. Obtaining E18-VP1 protein through prokaryotic expression; 2. Immunizing a C57BL/6 mouse to obtain E18-VP1 antiserum; 3. predicting and screening E18-VP1 protein epitope peptide; 4. c57BL/6 mice are immunized to obtain key epitope peptide antiserum; 5. And (3) detecting the neutralizing capacity of the key epitope peptide anti-serum virus. Specifically: The nucleotide sequence (OR 039189.1) of the Epstein-Barr virus 18-VP1 in the invention, the predicted amino acid sequence of the epitope peptide is shown as SEQ ID NO. 1-2, and the specific implementation flow is shown in figure 1. The invention firstly predicts and synthesizes the antigen epitope of E18-VP1 protein, screens the antigen epitope with binding capacity through E18-VP1 antiserum, immunizes mice with the screened key epitope peptide to prepare antiserum, and verifies the virus neutralization capacity of the serum. Specifically, E18 live strain is obtained from enterovirus infected clinical samples, VP1 nucleotide sequence is obtained through whole genome sequencing analysis, recombinant plasmid pET-28a (+) -E18-VP1 containing E18-VP1 gene sequence is constructed by taking pET-28a (+) as prokaryotic expression vector (see figure 2), E.coliBL21 (DE 3) is taken as expression strain to express E18-VP1 protein, target protein with purity of more than 90% is obtained after purification (see figure 3), C57BL/6 mice are immunized by taking E18-VP1 protein as immunogen, serum is collected and antibody titer is detected (see figure 4), epitope in E18-VP1 complete sequence is predicted through biological informatics analysis website, epitope exposed near FcRn is selected for synthesis, epitope capable of combining with E18-VP1 antiserum is screened through enzyme-linked immunosorbent assay, and epitope sequence from E18-VP1 protein is finally obtained (see figure 5, amino acid sequence is shown as sequence table SEQ ID NO: 1). The screened key epitope peptides were synthesized as immunogens after immunization of C57BL/6 mice, serum was collected and antibody titer was detected (see fig. 6), and virus neutralization ability was verified by qPCR (see fig. 7). Advantageous effects the epothilone genome comprises