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Anti-respiratory syncytial virus fully human broad-spectrum neutralizing antibody 4f1 and its application

A syncytial virus, anti-respiratory technology, applied in the field of medicine, can solve the problems of high price and high dosage

Active Publication Date: 2022-06-28
CENT FOR EXCELLENCE IN MOLECULAR CELL SCI CHINESE ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no vaccine or commercially available treatment for RSV in China, and only ribavirin is approved for the treatment of RSV infection, but has serious side effects
Palivizumab targets the conserved region of the F protein and has a broad spectrum of action. However, the current dose of Palivizumab required for each administration is 15 mg / kg (body weight), and passive immunization is performed 5 times a year. High, need multiple administrations, expensive disadvantages, and Palivizumab is a humanized antibody, which still retains part of the mouse sequence, may have certain immunogenicity, and has a certain degree of safety concern

Method used

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  • Anti-respiratory syncytial virus fully human broad-spectrum neutralizing antibody 4f1 and its application
  • Anti-respiratory syncytial virus fully human broad-spectrum neutralizing antibody 4f1 and its application
  • Anti-respiratory syncytial virus fully human broad-spectrum neutralizing antibody 4f1 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0193] Example 1 Obtaining antibody genes and antibody expression by single-cell RT-PCR method

[0194] 1. Acquisition of peripheral blood mononuclear cells (PBMC)

[0195] Peripheral blood was drawn from healthy volunteers, using conventional Ficoll-Paque (manufactured by -H (CEDARLANE) density gradient centrifugation to obtain 10 7 The above peripheral blood mononuclear cells (PBMC).

[0196] Ficoll separation method:

[0197] (1) Collect blood, collect 20ml of whole blood in a 50ml centrifuge tube (pre-containing 1ml of 4% sodium citrate), invert and mix 8-10 times. (even if the final concentration of sodium citrate is 0.4%);

[0198] (2) add an equal volume of RPMI1640 (containing sodium citrate), and mix;

[0199] (3) Using a 15ml transparent centrifuge tube, spread 3ml of lymphocyte separation solution, and carefully add 6ml of blood sample on it. Form a separation interface (or 4ml separation solution plus 8ml blood sample);

[0200] (4) Centrifuge 800g at room ...

Embodiment 2

[0226] Example 2 Antibody Characterization Analysis

[0227] 1. ELISA detects the activity of antibody binding to antigen

[0228] To investigate the ability of 4F1 antibody to bind to RSV type A and B virus F proteins, ELISA was used to detect whether the expressed antibodies recognized RSV A2 and B9320 pre-fusion F proteins and A2 post-fusion F proteins. The A2F protein sequence is referred to UniProtKB / Swiss-Prot: P03420.1, and the B9320 sequence is referred to UniProtKB / Swiss-Prot: Q6V2E7. The design of RSV pre-fusion F protein is based on the strategy adopted by Jason S. McLellan. Science 2013. RSV post-fusion F The protein was designed according to the strategy adopted in Davide Corti.Nature 2013. The whole gene was synthesized in Shanghai Jierui Company and constructed into the expression vector of invitrogen pcDNA3.1. Expressed by mammalian cell CHO expression system; refer to Invitrogen ExpiCHO-S TM Expression System Handbook. Palivizumab (Palivizumab, ) is the p...

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Abstract

The invention discloses a fully human neutralizing antibody against fusion protein of respiratory syncytial virus and application thereof. Specifically, the present invention discloses a fully human monoclonal antibody 4F1 against the fusion protein (F protein) and pre-fusion protein (preF protein) of respiratory syncytial virus, nucleic acid sequences encoding the antibody and antibody fragments, and a preparation method thereof. In vitro and in vivo experiments have confirmed that the 4F1 antibody can effectively prevent and control RSV infection. It has low immunogenicity for the human body and can avoid human-anti-mouse and other antibody-mediated immune rejection reactions. It can be used clinically For the prevention and treatment of respiratory syncytial virus infection.

Description

technical field [0001] The invention relates to the field of medicine, in particular to a fully human broad-spectrum neutralizing antibody 4F1 against respiratory syncytial virus and its application. Background technique [0002] RSV causes acute upper and lower respiratory tract infections and is one of the most important pathogens of respiratory tract infections in infants and young children worldwide. At the same time, RSV is also recognized as a high-risk adult, such as elderly individuals, adults with chronic lung disease Important pathogens in individuals as well as in immunocompromised adults such as bone marrow transplant patients. There are 30 million new infections worldwide each year, and about 200,000 people die from RSV infection. Among Chinese children under 2 years of age with pneumonia syndrome, RSV was the most common viral pathogen (17.0%). There is an urgent need worldwide for safe, effective, and inexpensive preventive treatments for RSV virus infection...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/10A61K39/42A61P31/14G01N33/577G01N33/569
CPCC07K16/1027A61P31/14C07K2317/565C07K2317/56C07K2317/76C07K2317/92A61K2039/505C07K16/10A61K39/42G01N33/569G01N33/577A61K39/395C07K19/00
Inventor 孙兵王宾凌志洋赵干伊春艳
Owner CENT FOR EXCELLENCE IN MOLECULAR CELL SCI CHINESE ACAD OF SCI
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