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New HPV epitope

A molecular and sequence technology, applied in the field of novel HPV antigen epitopes, can solve the problem that the antigen epitope has not been reported, and achieve the effect of high detection positive rate

Inactive Publication Date: 2018-06-15
CANCER INST & HOSPITAL CHINESE ACADEMY OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the epitopes of hr-HPV oncoproteins E6 and E7

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1: Obtaining of HPV16 Oncoprotein E6 and E7 Epitopes

[0055] 1. Preparation of nitrocellulose membrane

[0056] Using spot synthesis (SPOT synthesis) technology (see, for example, Chinese Patent Application Publication No. CN1602204) to synthesize a peptide array covering the entire amino acid sequence of HPV16E6 / E7, each peptide segment is 18 amino acids in length, and the interval between two adjacent peptide segments The overlap length is 16 amino acids. After inputting the peptide synthesis sequence into the computer, the peptide synthesizer AutoSpot (available for example from Intavis AG, Germany) spots specific amino acids on the surface of the activated nitrocellulose membrane under the control of the MultiPep software program.

[0057] 2. Detect the presence of antibodies in the mixed serum of patients with esophageal squamous cell carcinoma

[0058] Soak the synthetic HPV16E6 / E7 polypeptide nitrocellulose membrane in 100%, 75% and 50% ethanol for 5 m...

Embodiment 2

[0069] Example 2: Validation of antigenic epitopes in large population samples

[0070]Spot the obtained epitope sequence on the iPDMS polypeptide chip, and the chip is hybridized with serum samples from different individuals. Wet the surface of the peptide chip reactor with serum diluent for 3 minutes; use serum diluent (1% bovine serum albumin, 1% casein, 0.5% sucrose, 0.2% polyvinylpyrrolidone, 0.5% Tween-20, 0.01 M phosphate buffer, pH=7.4) was diluted to 200 μL at 1:200, transferred to the peptide chip reactor and incubated on a shaker (4°C, 120 rpm, incubation for 2 hours), in each batch of experiments should be Set up a negative control (200 μl serum diluent); rinse the surface of the chip with washing solution for 3 times after incubation, and blot the remaining washing solution with an aspirator; add goat anti-human-IgG to each reactor, and incubate statically (4 ℃, 1 hour), then rinse with lotion for 3 times; dry the surface of the chip with an ear wash ball, mix A ...

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Abstract

The present invention relates to new HPV epitope, wherein the sequence of the new HPV epitope is an amino acid sequence selected from SEQ ID NO:1-5 or an amino acid sequence having at least 80% identity with a sequence selected from SEQ ID NO:1-5. The invention further relates to applications of the epitope in detection of the presence of antibodies against HPV, particularly high-risk HPV, in samples, and in detection of the infection of HPV, particularly high-risk HPV, in subjects.

Description

field of invention [0001] The present invention relates to a novel HPV antigenic epitope and its application. Background of the invention [0002] Esophageal cancer is the eighth most common cancer in the world. In China, the main histological type of esophageal cancer is esophageal squamous cell carcinoma (ESCC). The occurrence of esophageal squamous cell carcinoma is the result of the interaction between environmental factors and genetic factors. Environmental factors include eating habits, smoking, drinking, etc., and may be related to viral infections. [0003] Infection is one of the factors of tumorigenesis. In 2008, tumors caused by infection accounted for 16.1% of all tumors. Among them, human papillomavirus (Human Papillomavirus, HPV) is an important carcinogenic virus. It is reported that 90% of cervical squamous cell carcinomas are related to HPV infection, and there is also a subtype related to HPV infection in oropharyngeal squamous cell carcinoma. In additi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/025C07K16/08C12N15/37G01N33/569A61K39/12A61P31/20A61P35/00
CPCA61K39/00C07K14/005C07K16/084C12N2710/20022G01N33/56983
Inventor 赵晓航周典蓉郑巍薇孙玉琳代淑阳刘芳周兰萍
Owner CANCER INST & HOSPITAL CHINESE ACADEMY OF MEDICAL SCI
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