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Universal Ebola virus disease immunoglobulin and its preparation method and application

A technology of Ebola virus and protein, applied in the direction of antiviral immunoglobulin, immunoglobulin, immunoglobulin from serum, etc.

Active Publication Date: 2021-11-26
ACAD OF MILITARY SCI PLA CHINA ACAD OF MILITARY MEDICAL SCI INST OF MILITARY VETERINARY MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, there is no effective preventive and curative drug for the disease

Method used

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preparation example Construction

[0044] The present invention provides a safe, efficient and reliable refined universal Ebola virus disease immunoglobulin and a preparation method thereof, in particular a complete set of immunogens for producing and refined universal Ebola virus disease immunoglobulin.

[0045] The complete set of immunogens for purifying universal Ebola virus disease immunoglobulin provided by the present invention is composed of Ebola virus virus-like particle vaccine antigens and / or vaccinia virus recombinant live vector vaccine antigens.

[0046] First, analyze the genomes and antigenic epitopes of Zaire, Sudan, and Cote d'Ivoire Ebola viruses, design and synthesize the GP and VP40 genes of Ebola virus and optimize cell-preferred codons; second, use insect-rod The virus expression system and / or the vaccinia virus recombinant live vector vaccine system expresses, purifies and prepares the chimeric Ebola virus-like particle vaccine antigen; again, immunize healthy animals to obtain highly im...

Embodiment 1

[0062] Example 1 Horse anti-Ebola virus immunoglobulin F (ab') 2 and preparation method thereof

[0063] Equine anti-Ebola virus immunoglobulin F(ab') 2 The preparation method comprises the following steps:

[0064] 1. Design and synthesis of Ebola virus GP and VP40: Download the Ebola virus sequence from NCBI, and use bioinformatics software such as DNAMAN to analyze biological information such as gene homology, antigenic sites, and preferred codons; according to Ebola virus The structural characteristics of the virus were improved, and the GP and VP40 gene sequences of Ebola virus were optimized to fully cover the main antigenic epitopes of Zaire, Sudan and Cote d’Ivoire Ebola viruses, and the cell-preferred codons were optimized according to the expression system. The GP nucleotide sequence is shown in SEQ ID NO:1, and the VP40 nucleotide sequence is shown in SEQ ID NO:2.

[0065] 2. Preparation of VLP protein immunogen:

[0066] The two Ebola virus genes, GP and VP40, ...

Embodiment 2

[0070] Embodiment 2 Ebola virus disease purified immunoglobulin and its preparation method

[0071] The preparation method of Ebola virus disease purified immunoglobulin comprises the following steps:

[0072] 1. Design and synthesis of Ebola virus GP and VP40: Download the Ebola virus sequence from NCBI, and use bioinformatics software such as DNAMAN to analyze biological information such as gene homology, antigenic sites, and preferred codons; according to Ebola virus The structural characteristics of the virus were improved, and the GP and VP40 gene sequences of Ebola virus were optimized to fully cover the main antigenic epitopes of Zaire, Sudan and Cote d’Ivoire Ebola viruses, and the cell-preferred codons were optimized according to the expression system. The nucleotide sequence of GP is shown in SEQ ID NO:1, and the nucleotide sequence of VP40 is shown in SEQ ID NO:2.

[0073] 2. Preparation of immunogen:

[0074] The transfer plasmid was constructed based on the uppe...

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Abstract

The invention provides a general Ebola virus disease immunoglobulin and its preparation method and application. The present invention firstly analyzes the Zaire, Sudan and Côte d'Ivoire Ebola virus genomes, optimizes the Ebola virus GP and VP40 gene sequences, and forms a "chimeric virus-like particle" sequence so that it can fully cover the three types of viruses The main epitope of the Ebola chimeric virus-like particle vaccine was expressed, purified and prepared using the insect-baculovirus expression system and the vaccinia virus recombinant live vector vaccine system respectively; finally, the hyperimmune plasma was obtained from the immunized animals, High-purity anti-Ebola virus disease immunoglobulin F(ab') was obtained by digestion with pepsin, precipitation with ammonium sulfate, and column chromatography 2 , and evaluate its safety and efficacy. The universal Ebola virus disease immunoglobulin provided by the invention can prevent and treat hemorrhagic fever caused by three types of Ebola virus infection, and provide a powerful tool for the prevention and control of Ebola epidemics.

Description

technical field [0001] The invention relates to the field of biopharmaceuticals, in particular to a general Ebola virus disease immunoglobulin and a preparation method and application thereof. Background technique [0002] Ebola hemorrhagic fever is caused by Ebola virus (EBOV), with a fatality rate of up to 90%. Since the disease was discovered in the Ebola River Basin in Africa in 1976, it has spread every year. The WHO has recognized it as a global public health emergency and called on the world to strengthen drug research and development. [0003] EBOV includes five subtypes, Ebola-Zaire (Z-EBOV), Ebola-Sudan (S-EBOV), Ebola-Cote d'Ivoire (Ebola-Coted'Ivoire) , C-EBOV), Ebola-Reston (R-EBOV) and Ebola-bundibugyo (B-EBOLA). Among them, Z-EBOV is the most virulent, and has the highest mortality rate after infection in humans; S-EBOV is the second; C-EBOV is pathogenic to chimpanzees, but the virulence is the weakest to humans; R and B-EBOV are not effective to humans Pa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/40C07K14/08C12N15/866C12N15/863A61K39/12A61K39/42A61P31/14C07K16/10C07K16/06
CPCA61K39/12A61K2039/505A61K2039/5256A61K2039/5258A61P31/14C07K14/005C07K16/06C07K16/10C07K2317/33C07K2317/54C07K2317/76C07K2317/94C12N15/86C12N2710/14043C12N2710/24043C12N2710/24243C12N2760/14022C12N2760/14034C12N2760/14052C12N2800/105C12N2800/30
Inventor 赵永坤赵忠鹏张颖杨松涛高玉伟闫飞虎郑学星王化磊李敏杨晓岚夏咸柱
Owner ACAD OF MILITARY SCI PLA CHINA ACAD OF MILITARY MEDICAL SCI INST OF MILITARY VETERINARY MEDICINE
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