Anti-HPV (human papilloma virus) antibody and preparation method and application thereof

A HPV16 and antibody technology, applied in the field of immunotherapy, can solve the problems of low complex efficiency, interference of target monoclonal antibody screening and epitope identification, cumbersome method process, etc., and achieve high specificity effect

Active Publication Date: 2012-05-09
SHANGHAI ZERUN BIOTECHNOLOGY CO LTD
View PDF3 Cites 9 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The method using MHC molecules needs to express MHC molecules in vitro and form complexes between MHC molecules and antigenic peptides through in vitro renaturation. This method is cumbersome and the efficiency of preparing complexes is low.
The use of macromolecular coupling such as KLH also faces the problem of low efficiency in preparing complexes
Moreover, because the MHC or KLH molecules that have not formed complexes are also immunogenic, many unwanted interfering antibodies will be produced after immunization into the animal body, which will bring great difficulties to the subsequent target monoclonal antibody screening and epitope identification. interference

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
  • Anti-HPV (human papilloma virus) antibody and preparation method and application thereof
  • Anti-HPV (human papilloma virus) antibody and preparation method and application thereof
  • Anti-HPV (human papilloma virus) antibody and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0049] The method for preparing antibodies of the present invention may further include purifying the collected antibodies.

[0050] The present invention also provides a cell that produces an antibody of the present invention. In one embodiment, it is a 6C10 cell preserved in the General Microorganism Center of China Committee for Culture Collection of Microorganisms (CGMCC) with a preservation number of CGMCC 4144.

[0051] Purification of the antibodies of the present invention can be performed by conventional purification methods known to those of ordinary skill in the art, such as affinity chromatography, ion exchange chromatography, hydrophobic chromatography, molecular sieve chromatography, and the like.

[0052] The antibody of the present invention can be used to prevent or treat HPV16-infected diseases, including administering an effective amount of the antibody of the present invention to a subject, and a particularly preferred antibody is a monoclonal antibody produ...

Embodiment 1

[0055] Example 1: Preparation and immunization of immune complexes of E7 protein 49-57 peptide

[0056] 49-57 peptide of HPV16 virus E7 protein, amino acid sequence: RAHYNIVTF; molecular weight 1119Da, theoretical isoelectric point 9.04. The peptide was synthesized by Jill Biochemical Company with a purity greater than 95%. Synthetic double-stranded RNA——PolyIC was used as an adjuvant, which was purchased from Sigma (Cat. No.: P9582).

Embodiment 2

[0058] Embodiment 2: the detection of serum antibody titer

[0059] For the mice immunized as described in Example 1, a small amount of serum was collected from the tail vein on the 35th day after the initial immunization, and the serum antibody titer was detected by direct enzyme-linked immunoassay (ELISA).

[0060] The steps for detecting serum antibody titer by ELISA are as follows:

[0061] 1. First use the HPV16E7 protein (the preparation method of the protein is: synthesize the full-length HPV 16E7 gene, then express the E7 fusion protein with the Trx tag in the pET32a vector and BL21(DE3) host, and then use enterokinase to remove the Trx tag HPV16E7 protein can be obtained by dropping), and the amount of 1 μg per well was coated on a 96-well plate. Incubate the coated plate at 4°C for 16 hours, wash each well 6 times with washing buffer (PBS solution containing 0.05% Tween-20), add 100 μl of blocking solution (PBS solution containing 5% skimmed milk powder) to each wel...

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

No PUM Login to view more

Abstract

The invention relates to an antibody used for treatment of diseases, in particular to cervical cancer, related to human papilloma virus infection, and a preparation method and an application thereof.

Description

technical field [0001] The present invention relates to the field of immunotherapy. More specifically, the present invention relates to an antibody for preventing or treating human papillomavirus (human papillomavirus, HPV) infection-related diseases (such as cervical lesions, especially cervical cancer). The present invention also relates to the preparation method and application of the antibody. Background technique [0002] Current research shows that HPV infection is significantly related to the occurrence of cervical system diseases, especially cervical cancer. In 1995, the research results released by the International Agency for Research of Cancer (IARC) confirmed that HPV has a close causal relationship with cervical cancer and other cervical system diseases: HPV can be detected in 99.8% of cervical cancer patients Infection, while HPV-negative patients will hardly develop cervical cancer; in addition, HPV can be detected in more than 98% of cervical disease patien...

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 Applications(China)
IPC IPC(8): C07K16/08C12N5/20A61K39/42A61P31/20A61P15/00A61P35/00C12R1/91
Inventor 雷建强丛薇田平生刘瑞锋张高峡
Owner SHANGHAI ZERUN BIOTECHNOLOGY CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap