Monoclonal neutralizing antibody against hpv58l1 and its application

A monoclonal antibody and protein technology, applied in the fields of antibodies, applications, anti-tumor drugs, etc., can solve the problems of less research on monoclonal antibodies, restrictions on monoclonal antibody detection of HPV58 virus HPV58 virus treatment, etc., and achieve the goal of improving the clinical negative conversion rate Effect

Active Publication Date: 2022-05-20
中生方政生物技术股份有限公司
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] It can be seen from the above data that HPV58 is a common high-risk type after HPV16 and HPV52 in HPV infection cases and cervical cancer patients, but there are very few researches on its monoclonal antibody, which limits the use of monoclonal antibody detection HPV58 virus, and the treatment of HPV58 virus

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
  • Monoclonal neutralizing antibody against hpv58l1 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0098] Embodiment 1, establishment of hybridoma cells

[0099] 1. Animal immunization

[0100] Using the Escherichia coli expression system, the virus packaging particles of the L1 protein of HPV58 type were prepared as the antigen. For the first immunization, the antigen was mixed with Freund's complete adjuvant in equal volumes and fully emulsified, and injected subcutaneously in points. Each Balb / c mouse The injection volume was 100 μg, and 3 mice were immunized; on the 7th, 14th, and 21st days of the first immunization, the emulsion of antigen and Freund's incomplete adjuvant was used for booster immunization, and on the 14th day, small Blood was collected from the tail vein of the rat, the serum was separated, and the antibody titer was detected by indirect ELISA method. The antibody titer of the mouse serum was 1:16000, >1:32000, and 1:16000 respectively, and the No. 2 mouse with the highest titer was selected for fusion .

[0101] The operation steps of the indirect E...

Embodiment 2

[0110] Example 2: Preparation and identification of monoclonal antibodies

[0111] 1. Preparation of anti-HPV58 monoclonal antibody from mouse ascites

[0112] Adult BALB / c mice were selected, and hybridoma cells 58-4A3 were inoculated intraperitoneally, 1×10 per mouse 6 -2×10 6 First, when the abdomen is obviously enlarged and the skin feels tense when touched with hands, ascites can be collected with a 16-gauge needle. Centrifuge the ascites (13000r / min for 30 minutes), remove cell components and other precipitates, collect the supernatant, purify the antibody with a ProteinG affinity column, and detect the titer of the ascites and the purified antibody by indirect ELISA. The results show that the ascites was diluted 100,000 times positive, the purified antibody diluted to 1ng / mL is still positive. Antibody Purity Determination

[0113] 2. Perform 12% SDS-PAGE electrophoresis on the purified antibody, the result shows that the purity is above 95%, see figure 1 As shown,...

Embodiment 3

[0119] Example 3: Detection of antibody neutralization activity

[0120] Through the pseudovirus-cell neutralization model, the neutralizing activity of the antibody was tested.

[0121]First dilute the antibody to 1000ng / mL with PBS, then dilute the antibody to 0.06ng / mL by 2-fold gradient, take 50μL of each concentration of antibody and 50μL of appropriate concentration of HPV6, 11, 16, 18, 31, 33, 45, 52, 58, 59, and 68 pseudoviruses were incubated in a 96-well plate at 4°C for one hour, and a negative serum control, a positive serum control, a cell control, and a pseudovirus control were set. Then each mixture was added to the 96-well plate pre-coated with 293FT cells and cultured in a cell culture incubator for 72 hours. Afterwards, the fluorescence was observed, and the cells were collected to detect the fluorescence by flow cytometry. If there was an inhibitory effect, the fluorescence inhibition rate was calculated according to the fluorescence inhibition rate = (1-ex...

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 the technical field of antibody medicine, in particular to a monoclonal neutralizing antibody of HPV58L1 and its application. The invention provides a specific monoclonal neutralizing antibody of HPV58 and a hybridoma cell line producing the antibody, using one of the monoclonal neutralizing antibodies with the highest neutralizing activity titer, using indirect ELISA method, double antibody sandwich ELISA method, immune hybridization method, etc., can be widely used in the detection of HPV58 virus infection in clinical samples; use one of the monoclonal neutralizing antibodies with the highest neutralizing activity to prepare vaginal gel, vaginal washing liquid, external use Lotion, vaginal suppository, freeze-dried powder, vaginal effervescent tablet, etc., are used for the treatment of patients with HPV58 infection, which can effectively improve the clinical negative conversion rate of HPV58 patients. The invention has important significance for women's healthy development and public health prevention and control.

Description

technical field [0001] The invention relates to the technical field of antibody medicine, in particular to a monoclonal neutralizing antibody of HPV58L1 and its application. Background technique [0002] Human papillomavirus (HPV) is a spherical, double-stranded DNA virus with a diameter of 55-60nm and a nucleocapsid with 20-hedral symmetry. Capsid protein L2 composition. It mainly invades human epithelial tissues and cells, can cause squamous epithelial proliferation of human skin and mucous membranes, and induce various benign and malignant hyperplastic lesions. High-risk HPV infection is associated with the occurrence of various malignant tumors, while low-risk HPV infection can cause anal and genital warts. [0003] High-risk HPV types include 16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 73, 82, etc., which can cause cervical intraepithelial neoplasia ( CIN) and cervical cancer, wherein HPV16 and HPV18 are the most common carcinogenic types, and the...

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): C07K16/08C12N15/13G01N33/68G01N33/577G01N33/569A61K39/42A61P31/20A61P35/00A61P17/12
CPCC07K16/084G01N33/68G01N33/577G01N33/56983A61P31/20A61P35/00A61P17/12C07K2317/565G01N2333/025A61K2039/505
Inventor 魏颖颖邹国宝刘欣欣郭光华蔺皓宋高尚沈江卫
Owner 中生方政生物技术股份有限公司
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