Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Human papilloma virus (HPV) capsid protein L1 polypeptide and preparation and application thereof

A capsid protein, papilloma technology, applied in the field of peptides

Inactive Publication Date: 2012-11-21
SICHUAN UNIV
View PDF3 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it often leads to overtreatment and related complications, and increases the psychological and economic burden of patients. Clinically, new detection methods are urgently needed to divert low-risk and high-risk patients.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Human papilloma virus (HPV) capsid protein L1 polypeptide and preparation and application thereof
  • Human papilloma virus (HPV) capsid protein L1 polypeptide and preparation and application thereof
  • Human papilloma virus (HPV) capsid protein L1 polypeptide and preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] In the primary immunization group, complete Freund's adjuvant and synthetic immunogen Cys Thr Leu Thr Ala Asp Val

[0039] Met Thr Tyr Ile His-KLH proteins were mixed in equal amounts and stirred on a magnetic stirrer for 30 minutes. After mixing evenly, rabbits were immunized, and each rabbit was immunized with 0.5ml of 200ug / ml antigen. Two weeks later, the same amount of immunogen was mixed with Freund's incomplete adjuvant, fully emulsified, and rabbits were immunized (the operation was the same as before). Immunize once every 2 weeks after the immunization, and immunize 4 times in total. Serum was collected from the carotid artery catheter 2 weeks after the fourth immunization, added with an equal amount of glycerol, mixed thoroughly and distributed.

[0040] Implementation example two

[0041] Immunocytochemical detection of the reactivity of anti-polypeptide antiserum to cells infected by HPV virus: HeLa (cervical adenocarcinoma cell line infected with HPV18), ...

Embodiment 2

[0044]Immunohistochemical detection of the reactivity of anti-polypeptide antiserum to HPV-positive clinical specimens: tissue specimens identified as condyloma acuminata, CIN, cervicitis and HPV-infected positive by the clinical pathology department were fixed in 10% formaldehyde, embedded in paraffin, sliced, After deparaffinization and hydration, repair with 0.01mol / lpH6.0 citrate buffer at 100°C for 20min, cool to room temperature naturally, wash with PBS for 5min×3 times, 3%H 2 o 2 Eliminate endogenous peroxidase for 30 min, PBS 5 min x 3 times, block with goat serum blocking solution for 30 min, add 1:500 diluted rabbit serum overnight at 4°C, wash with PBS for 5 min x 3 times, then add goat anti-rabbit II dropwise Anti-working solution for 15 minutes, wash with PBS for 5 minutes×3 times, add horseradish-labeled streptavidin working solution, wash with PBS for 5 minutes×3 times, develop color with DAB for 3 minutes, stop the reaction with tap water, counterstain with hem...

Embodiment 3

[0047] Western Blot detection of the reactivity of anti-peptide antiserum to cells infected with HPV virus:

[0048] A. Extraction of cell protein Total cell protein was extracted with BCA total protein extraction kit. Scrape off Siha (cervical squamous cell line infected with HPV16) and Hela (cervical adenocarcinoma cell line infected with HPV18) cells with a curette, centrifuge at 500g for 2min at 4°C, wash twice with PBS, add 200ul of RIPA, 8ul of cocktail, and 2ul of PMSF , 2ul protease inhibitors, shake for 15s, let stand on ice for 10min, centrifuge at 15000g 4°C for 10min, wash the supernatant and get the total protein. The protein concentration was determined with the BCA protein quantification kit, and stored in a -80°C refrigerator for later use.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
diameteraaaaaaaaaa
Login to View More

Abstract

The invention relates to a human papilloma virus capsid protein L1 (HPVL1) peptide and preparation and application thereof. The HPVL1 peptide is a peptide segment of which the amino acid of the sequence is N end-Cys Thr Leu Thr Ala Asp Val Met listed Thr Tyr Ile His-C end, or is a peptide segment comprising the amino acid of the sequence and with a length less than 50 amino acids. In the invention, the peptide which is a very conservative peptide segment located on the surface of protein is sieved out by performing epitope forecasting and multi-sequencing comparison on the L1 proteins of 18 low-risk types and high-risk type HPV subtypes (6, 11, 16, 18, 52, 58 and the like); the antibody generated from induction can react with multiple types of HPVL1, and can be used for the detection of the multiple types of HPVL1, in particular to the application of precancerous lesions detection of human cervical carcinoma; and the antibody aiming at the peptide segment also can be used for purifying and preparing the multiple types of HPVL1. In addition, the invention also provides the applications of the peptide segment used as multiple types of HPV antiserum and detected matters of monoclonalantibody.

Description

Technical field: [0001] The invention relates to a polypeptide, in particular to a human papilloma virus (HPV) capsid protein L1 polypeptide and its preparation and application. technical background: [0002] Cervical cancer is a disease that seriously affects women's health. Globally, cervical cancer is the second most common cancer among women, and even ranks first in some developing countries, with nearly 500,000 new cases each year (Valdespino VM, Valdespino VE..Curr Op in Obstet Gynecol, 2006, 18: 35- 40.). In recent years, human papillomavirus (HPV) infection is considered to be the main cause of cervical cancer. In 1999, Warlboomers et al. found that the detection rate of human papillomavirus (Human papillomavirus, HPV) was 99.7% in the study of 1035 cases of cervical cancer specimens in 22 countries. Since the first case of human papillomavirus (HPV) was discovered in 1933, more than 200 subtypes of human papillomavirus have been identified so far. According to t...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/08C07K16/08C07K16/06G01N33/577G01N33/569C12N7/02
Inventor 胡丽娜王霞刘宝林勇陈恒禧张林甘晓玲
Owner SICHUAN UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com