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Antigen fragment and truncated protamine based on Marburg virus envelope protein and application

A Marburg virus and protein technology, applied in the direction of virus antigen components, antiviral immunoglobulin, virus/bacteriophage, etc.

Active Publication Date: 2017-08-08
INST OF BIOENG ACAD OF MILITARY MEDICAL SCI OF THE CHINESE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, there are no effective vaccines and drugs for the prevention and treatment of Marburg virus infection

Method used

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  • Antigen fragment and truncated protamine based on Marburg virus envelope protein and application
  • Antigen fragment and truncated protamine based on Marburg virus envelope protein and application
  • Antigen fragment and truncated protamine based on Marburg virus envelope protein and application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0084] Embodiment 1, the preparation of antigen and fragment thereof

[0085] 1. Construction of recombinant plasmids

[0086] At the same time, the following recombinant plasmids were constructed: pVAX1-GP, pVAX1-GP△, pVAX1-RBS, pVAX1-RBSP and pVAX1-FU. Specific steps are as follows:

[0087] (1) Design and synthesis of primers

[0088] Primers used to construct recombinant plasmids pVAX1-GP, pVAX1-GP△, pVAX1-RBS, pVAX1-RBSP and pVAX1-FU were designed, and the primer sequences are shown in Table 1.

[0089] Table 1. Primer sequences for constructing recombinant plasmids

[0090] GPF 5’-CCGGAATTCGCCACCATGAAAACTACTTGCTTATTAAT GPR 5'-CCGCTCGAGCTTGTCATCGTCGTCCTTGTAGTCTTAGCCGATGTATTTAGTGAA GP△F 5'-CCGGAATTCGCCACCATGCTGCCAATCCTGGAGATCGC GP△R 5'-CCGCTCGAGTTATTACTTGTCATCGTCGTCCTTGTAGTCATCAGAAGTCCACCACTTGC RBSF 5’-CCGGAATTCGCCACCATGGCTTCTAATATTCAGCCACA RBSR 5'-CCGCTCGAGTTATTACTTGTCATCGTCGTCCTTGTAGTCGTGTCTGTAGCCCTGACC RBSP+ 5'-CC...

Embodiment 2

[0132] Embodiment 2, the immune activity assay of antigen and fragment thereof

[0133] (1) Immunization method

[0134] 1. Grouping

[0135] Animals Selected 6-week-old Balb / C female mice and divided them into normal saline group 1, pVAX1-GP△ group, pVAX1-RBS group, pVAX1-RBSP group, pVAX1-FU group, normal saline group 2, BSA protein group, GP1 △ protein group, GPM protein group, GP2△ protein group and GP mixed protein group (GP1△ protein, GP2△ protein and GPM protein were mixed at a mass ratio of 1:1:1), with 12 mice in each group.

[0136] 2. Immunization method

[0137] Normal saline 1 group, pVAX1-GP△ group, pVAX1-RBS group, pVAX1-RBSP group, and pVAX1-FU group were injected into the hind legs without adjuvant, and immunized once every two weeks (on day 0, day 14 and Immunization on the 28th day), 100 μg plasmid / only (plasmid concentration 1mg / ml), the immunization volume of physiological saline is 100 μl / only, totally 3 times, inject 0.25% procaine hydrochloride to st...

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Abstract

The invention discloses an antigen fragment and a truncated protamine based on Marburg virus envelope protein and application. The antigen fragment GP2 provided by the invention has antigen potential similar to GP protein and a neutralizing antibody generated by immunization can be used for effectively inhibiting infection of viruses. Compared with a complete GP protein antigen, the antigen fragment does not have a biological function of the GP protein, so that the viruses cannot enter host cells; when the segment is used for constructing virus-vectored vaccines, the segment has better safety and is more convenient for preparation of multivalent vaccines.

Description

technical field [0001] The invention relates to an antigen fragment, a truncated body and an application based on a Marburg virus envelope protein, and belongs to the field of biotechnology. Background technique [0002] Marburg virus (MARV) belongs to the Filoviridae family. It is a single-stranded non-segmented negative-strand RNA virus that is filamentous under an electron microscope. It is the causative agent of the severe infectious disease Marburg hemorrhagic fever. The virus was discovered by German scientists in 1967 (Siegert R, Shu HL. Ger Med Mon, 1968, 13(1): 1–2.), and so far there have been more than ten sporadic outbreaks in Africa and other places, causing dozens of infections. One hundred people died, and the mortality rate was greater than 80%. The Marburg hemorrhagic fever outbreak in Angola in 2005 had a mortality rate as high as 90% (Jonathan S. Towner ML. Virol, 2006, 80(13): 6497-6516.). MARV infection can cause severe hemorrhagic fever, destroying var...

Claims

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Application Information

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IPC IPC(8): C07K14/08C12N15/40C07K16/10A61K39/12A61P31/14
CPCA61K39/12C07K14/005C07K16/10C12N2760/14222C12N2760/14233C12N2760/14234
Inventor 刘珠果戴秋云李拓
Owner INST OF BIOENG ACAD OF MILITARY MEDICAL SCI OF THE CHINESE
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