Horse anti-EBOV (Ebola virus) immune globulin F (ab')2 and preparation method thereof

An Ebola virus and immunoglobulin technology, applied in the field of equine anti-Ebola virus disease immunoglobulin F2 and its preparation, can solve problems such as endangering public health and safety, achieve product safety and reliability, improve survival rate, and satisfy effect of supply

Inactive Publication Date: 2016-01-20
MILITARY VETERINARY RES INST PLA MILITARY MEDICAL ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, the situation of international and domestic political struggles is becoming more and more complicated. The Ebola virus, which is included in the list of the Convention on the Prohibition of Biological Weapons, may also be used by terrorists and hostile forces, endangering my country's public health security
Currently, there is no effective preventive and curative drug for the disease

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1 horse anti-Ebola virus disease immunoglobulin F (ab') 2 and its preparation method

[0024] Equine anti-Ebola virus disease immunoglobulin F(ab’) 2 The preparation method comprises the following steps:

[0025]1. Preparation of immunogen: two genes GP and VP40 (GenBankAccessionNo.AY354458, Zaire1995) of Zaire type Ebola virus were cloned on the same insect baculovirus expression vector pFastBacDual, transformed into Escherichia coli DH10Bac competent cells, The positive plasmid extracted is the recombinant baculovirus Bacmid plasmid, and the recombinant baculovirus Bacmid plasmid is used to transfect the insect cell Sf9, and the recombinant baculovirus is rescued, and the obtained recombinant baculovirus is inoculated into the insect cell Sf9, and cultured at 27°C for 48- After 72 hours, the cell supernatant was harvested, and virus-like particles expressing both GP and VP40 genes of Zaire-type Ebola virus were obtained.

[0026] After identification, th...

Embodiment 2

[0040] Embodiment 2 horse anti-Ebola virus disease immunoglobulin F (ab') 2 Treatment Effect Evaluation

[0041] 4-6 weeks old female BALB / c mice (15-19 g) were randomly divided into 3 groups, 8 mice in each group. All mice were inoculated with 1000 times LD by intraperitoneal injection 50 Doses of murine-adapted Ebola virus (MA-EBOV, USAMRIID / BALB / c-lab / COD / 1976 / Mayinga-MA-p3, provided by the U.S. Army Medical Research Institute of Infectious Diseases) at 1 and 2 days post-infection 200 μg F(ab') by intraperitoneal injection 2 Infected mice were treated at a dose of / only / day, twice a day, for 3 consecutive days, and mice in the control group were treated with the same volume of PBS 1 day after virus infection. The mice were weighed every day and the state of the mice was observed for 16 consecutive days, and the surviving mice were observed for another 12 days. The results showed that the mice in the control group lost about 23% of their body weight and all died with an ...

Embodiment 3

[0042] Embodiment 3 horse anti-Ebola virus disease immunoglobulin F (ab') 2 Treatment Effect Evaluation

[0043] 4-6 weeks old female BALB / c mice (15-19 grams) were randomly divided into 3 groups, 9-10 mice / group. All mice were inoculated with 1000 times LD by intraperitoneal injection 50 Dosage of murine adapted Ebola virus (MA-EBOV). 1 mg F(ab') by intraperitoneal injection 1 day after infection 2 / only / day and 2 mg F(ab') 2 Infected mice were treated at a dose of / only / day, twice a day, for 3 consecutive days, and mice in the control group were treated with the same volume of PBS 1 day after virus infection. The mice were weighed every day and the state of the mice was observed for 16 consecutive days, and the surviving mice were observed for another 12 days. The results showed that the mice in the control group lost about 17.3% of their body weight and all died with an average death time of 6.3±1.0 days. Mice were treated 1 day after viral infection with a therape...

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Abstract

The invention provides horse anti-EBOV (Ebola virus) immune globulin F (ab')2 and a preparation method thereof. The preparation method comprises steps as follows: virus-like particles of a GP gene and a VP40 gene expressing EBOV-Z simultaneously are inactivated and purified and then are mixed with an immunologic adjuvant to be taken as immunogens, highly immunized plasma is obtained through repeated immunization of healthy horses, IgG protein is separated and purified, then the IgG protein is subjected to pepsase digestion, an enzyme-digested product is processed by a DEAE ion-exchange chromatographic column, an eluent is collected and subjected to desalination and freeze-drying, and the horse anti-EBOV immune globulin F (ab')2 is obtained. The horse anti-EBOV immune globulin F (ab')2 is an effective drug for preventing and treating Ebola hemorrhagic fever caused by EBOV and has remarkable curative effects for critical patients infected with the EBOV; the preparation method is controlled by adopting unique technological parameters and is suitable for industrial production, and the product is safe and reliable.

Description

technical field [0001] The invention relates to the fields of immunology and genetic engineering, in particular to a horse anti-Ebola virus disease immunoglobulin F (ab') 2 and its preparation method. Background technique [0002] Ebola hemorrhagic fever is caused by Ebola virus (EBOV), and the case fatality rate can be as high as 90%. It is the most terrifying and severe infectious disease in the world today. Since the disease was discovered in the Ebola River Basin in Africa in 1976, it has occurred every year. This year's epidemic is particularly serious. Since March 2014, it has continued to break out in Guinea, Liberia, and Sierra Leone in Africa. It has spread rapidly and the epidemic is almost out of control. , As of October 9, 2014, 3,750 people have been diagnosed and 1,876 people have died. WHO calls on the world to strengthen prevention and control. [0003] my country is within the scope of the Ebola natural epidemic area announced by the WHO. There are natural...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/10C07K16/06C07K1/18A61K39/42A61P31/14
Inventor 夏咸柱杨松涛赵永坤郑学星王化磊高玉伟金宏丽王铁成范泉水郑颖王承宇黄耕陈维金冯娜曹增国盖薇薇毕钰海闫飞虎李月涛王翀迟航
Owner MILITARY VETERINARY RES INST PLA MILITARY MEDICAL ACAD OF SCI
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