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A fully human neutralizing antibody against rabies virus

A rabies virus, fully human technology, applied in the fields of cellular immunology and molecular biology, can solve the problems of long production cycle, low antibody affinity and limited success rate, and achieve good neutralization effect, good specificity and affinity. high effect

Active Publication Date: 2016-04-20
CHANGCHUN BCHT BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, if the monoclonal antibody of mouse origin is used as a drug in the human body, it will cause severe human anti-mouse antibody (HAMA) reaction, so it cannot be used directly. It must be humanized, and the production cycle is long and the success rate is limited.
[0006] At present, the development of human-derived monoclonal antibodies mainly includes phage display technology, transgenic animals, memory B cell immortalization and other technologies. Among them, phage display technology can obtain a large antibody library with high throughput, and the antibody is easy to purify. However, the obtained antibodies often have low affinity and poor therapeutic effect; the obvious advantage of the transgenic mouse method is that it is easy to operate and easy to obtain antibodies, but the disadvantage is that there are restrictions on intellectual property rights and animal models, which make the cost of antibodies produced by this technology high; and The experimental procedures for the production of antibodies by the immortalization of memory B cells are relatively complicated, most of which are still in the basic research stage, and the technical means have not yet reached the level of stable production

Method used

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  • A fully human neutralizing antibody against rabies virus
  • A fully human neutralizing antibody against rabies virus

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Embodiment Construction

[0018] The preparation process of the fully human monoclonal anti-rabies virus neutralizing antibody TRN006 described in the embodiments of the present invention comprises the following steps:

[0019] 1. Lymphocyte Isolation and Single Memory B Cell Sorting

[0020] Collect 100ml of venous blood from two volunteers who have been injected with rabies vaccine (Novartis batch number: 1928) and have produced protective antibodies, put them in anticoagulant tubes containing heparin, and use density centrifugation to separate mononuclear cells (PBMCs) ), after cell counting, single B cells of CD3-, CD14-, CD16-, CD19+, CD27+ were sorted from PBMC by BDFACSria flow cytometer (BD Biosciences, San Jose, CA) into a 96-well PCR plate containing RT reaction system, Make each well contain one memory B cell, and freeze the PCR plate at -80°C for future use.

[0021] 2. Isolation of Antibody Variable Region Genes from Single B Cells by RT-PCR

[0022] 1) Synthesize the first strand of c...

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Abstract

The invention discloses a completely humanized neutralizing antibody for anti-rabies viruses. The amino acid sequence of a heavy chain of the antibody is shown in sequence table SEQ ID NO1, the amino acid sequence of a light chain of the antibody is shown in sequence table SEQ ID NO5, the complementary determining region (CDR) of a variable region of the heavy chain of the antibody is shown as follows: CDR1: SEQIDNO2, CDR2:SEQIDNO3 and CDR3: SEQIDNO4, and the complementary determining region (CDR) of a variable region of the light chain of the antibody is shown as follows: CDR1:SEQIDNO6, CDR2:SEQIDNO7, and CDR3: SEQIDNO8. The beneficial effects of the completely humanized neutralizing antibody are that: the completely humanized neutralizing antibody disclosed by the invention ahs the advantages of being completely humanized, good in specificity, high in affinity, good in neutralizing effect, and low in price, can be used as a biological engineering antibody drug to quickly establish immune protection force against rabies virus, can be used for passive immunotherapy of acute infection, and also can be used for preparing detection reagents for detecting the rabies virus, finding effective neutralization antigen epitope and developing recombinant proteins and subunit vaccines for the rabies virus.

Description

technical field [0001] The invention belongs to the field of cellular immunology and molecular biology, relates to a human monoclonal antibody, in particular to a fully human neutralizing antibody against rabies virus. Background technique [0002] Rabies is a zoonotic infectious disease caused by the rabies virus, with a fatality rate greater than 90%. The disease is widely distributed all over the world. my country is a high-incidence area of ​​rabies, ranking second in Asia (Asia is a high-incidence area in the world). [0003] Once the human body is bitten by a dog or other animal carrying rabies virus, the most effective way is to use anti-rabies virus neutralizing antibody or anti-viral serum as soon as possible to quickly activate passive immune protection, so as to quickly eliminate the invasion into the body The purpose of the rabies virus. [0004] There have been antiviral serum applications of animal or human origin: the earliest application was the antirabies ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/10C07K16/46C12N15/13C12N15/63A61K39/42A61P31/14G01N33/577G01N33/569
Inventor 李姝余小玲王月明廖化新
Owner CHANGCHUN BCHT BIOTECH
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