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An anti-Ebola-virus immunoglobulin F(ab')2 and a preparing method thereof

An Ebola virus, protein technology, applied in antiviral immunoglobulins, antiviral agents, antibodies, etc.

Inactive Publication Date: 2015-08-12
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] So far, there is no effective method for the prevention and treatment of EBOV in the world

Method used

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  • An anti-Ebola-virus immunoglobulin F(ab')2 and a preparing method thereof
  • An anti-Ebola-virus immunoglobulin F(ab')2 and a preparing method thereof
  • An anti-Ebola-virus immunoglobulin F(ab')2 and a preparing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] Embodiment 1, EBOV nucleic acid vaccine and anti-EBOV specific immunoglobulin F (ab') 2 preparation of

[0082] One, the preparation of EBOV nucleic acid vaccine

[0083] 1. Obtaining the target gene

[0084]Download the full gene sequence (2031bp) of EBOV G protein from the NCBI website, use DNAMAN software to perform sequence alignment, analyze the homology of nucleotide and amino acid sequences, and obtain the nucleotide sequence that can represent the genetic characteristics of modern epidemic strains; analyze the G Protein nucleotide sequence, commercial nucleic acid vaccine vector pVAX1 sequence endonuclease cleavage site; endonuclease (EcoR Ⅰ) sequence (GAATTC) and KozaK sequence (ACTATGG) are added upstream of the G protein nucleotide sequence, and downstream Stop codon sequence (TAA) and endonuclease (Xba Ⅰ) sequence (TCTAGA), commercial company whole gene synthesis EBOV nucleic acid vaccine target gene sequence: the target gene sequence of Zaire type EBOV G ...

Embodiment 2

[0133] Embodiment 2, EBOV subunit vaccine and anti-EBOV specific immunoglobulin F (ab') 2 preparation of

[0134] 1. Acquisition of EBOV subunit vaccine

[0135] 1. Obtaining the target gene

[0136] Optimize the nucleotide sequence that can represent the genetic characteristics of modern epidemic strains, so that the codons are suitable for expression in insect cells; analyze the G protein nucleotide sequence and the sequence of the commercial insect expression vector pFastBac1 (Invitrogen, Cat. No.: 10360-014) Dicer enzyme cleavage site; an endonuclease (EcoR Ⅰ) sequence (GAATTC) is added upstream of the G protein nucleotide sequence, and a polyhistidine tag sequence (CACCACCACCACCACCAC), a stop codon sequence (TAA) and an endogenous sequence are added downstream Dicer (Xba Ⅰ) sequence (TCTAGA), the target gene sequence of the commercial company's whole gene synthesis EBOV subunit vaccine: the target gene sequence of Zaire type EBOV G protein is shown in sequence 4 in the ...

Embodiment 3

[0162] Embodiment 3, EBOV VLP vaccine and anti-EBOV specific immunoglobulin F (ab') 2 preparation of

[0163] 1. Preparation of EBOV VLP vaccine

[0164] 1. Obtaining the target gene

[0165] Optimize the nucleotide sequence that can represent the genetic characteristics of modern epidemic strains, so that the codons are suitable for expression in insect cells; analyze the G protein nucleotide sequence, VP40 protein nucleotide sequence, commercial insect expression vector pFastBacDUAL (purchased from Invitrogen Company , Cat. No.: 10712-024) endonuclease cleavage site; an endonuclease (EcoR Ⅰ) sequence (GAATTC) is added upstream of the G protein nucleotide sequence, and a stop codon sequence (TAA) and endonuclease are added downstream (Xba Ⅰ) sequence (TCTAGA); the endonuclease (Xho Ⅰ) sequence (CTCGAG) is added upstream of the VP40 protein nucleotide sequence, and the stop codon sequence (TAA) and endonuclease (KpnI) sequence (GGTACC) are added downstream , the target gene...

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Abstract

An anti-Ebola-virus immunoglobulin F(ab')2 and a preparing method thereof are disclosed. Three trivalent EBOV vaccine antigens, namely an EBOV nucleic acid vaccine, an EBOV subunit vaccine and an EBOV VLP vaccine, are utilized to immunize a healthy animal to produce a high-valence EBOV antiserum, and an Fc segment capable of causing side effects is removed to prepare the anti-EBOV specific immunoglobulin F(ab')2. Tests prove that the anti-EBOV specific immunoglobulin F(ab')2 prepared by the method is high in neutralization valence, purity and safety and stable, meets requirements of biological products in China, can specifically neutralize EBOV, has specific treatment effects for EBOV infection, can effectively treat Ebola virus disease and is a therapeutic drug with high value.

Description

technical field [0001] The invention relates to an anti-Ebola virus immunoglobulin F (ab') 2 The preparation method thereof belongs to the field of biotechnology pharmacy. Background technique [0002] EBOV (Ebola virus, EBOV) is one of the most severe zoonotic diseases, often causing fever, nausea, vomiting, diarrhea, skin color changes, body aches, internal bleeding, external bleeding and other symptoms, and often due to stroke, Myocardial infarction, hypovolemic shock or multiple organ failure lead to patient death, and the mortality rate of human infection can reach as high as 90%. EBOV includes five subtypes, Ebola-Zaire (Z-EBOV), Ebola-Sudan (S-EBOV), Ebola-Coted'Ivoire , C-EBOV), Ebola-Reston (R-EBOV) and Ebola-bundibugyo (B-EBOLA). Among them, Z-EBOV is the most virulent, and the mortality rate after human infection is the highest; S-EBOV is second; C-EBOV is pathogenic to chimpanzees, but the virulence is the weakest to humans; R-EBOV is not pathogenic to humans ...

Claims

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Application Information

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IPC IPC(8): C07K16/10A61K39/42A61P31/14
Inventor 赵忠鹏范志和范铁炯罗德炎李敏段跃强杨晓岚杨鹏辉邢丽王希良史小月李晓罗湘初张志平孙九如
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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