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Preparation method for immunoglobulinlg of adenovirus anti-Fi,anti-Pb and anti-Hx

An immunoglobulin and adenovirus technology, applied in the field of virology and immunology, which can solve the problems of no curative effect and certain drugs, etc.

Inactive Publication Date: 2008-07-16
TONROL BIOLOGICAL PHARM CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] At present, the commonly used drugs for antiviral infection are generally chemical synthetic drugs, Chinese herbal medicines, and biological preparations such as interferon. Among these drugs, there is no drug that is effective against viruses and has a certain effect

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Preparation of antigens containing adenovirus fibrin, penton protein and hexon protein

[0027] Take the fertilized eggs that have been hatched for 10-20 days, draw the air chamber and the fetal position, and find a place with less blood vessels near the embryo. After disinfection with iodine and ethanol, drill a small hole in the air chamber, and then place the egg membrane Puncture a small slit in the middle and suck out the air in the air chamber to separate the chorioallantoic sag from the air chamber, and drip into ADV (1×10 7 TCID50 / ml) 0.2ml, seal the mouth with sterile tape and seal the stomata with melted paraffin. After inoculation, the chicken embryos are placed at 37°C and incubated for 7 days. After incubation, sterilize the eggshell with iodine and ethanol, remove the eggshell along the edge of the air chamber with sterile forceps, lift the chorioallantoic membrane, puncture it with a sterile capillary pipette, absorb the allantoic fluid and collect...

Embodiment 2

[0028] Example 2: Preparation of antigens containing adenovirus fibrin, penton protein and hexon protein

[0029] According to conventional cell culture, after Hep-2 cells have grown into slices, add ADV (1×10 7 TCID50 / ml) was inoculated into Hep-2 cells that had grown into a monolayer, and incubated at 33°C. The cytopathic condition was observed every day. When the cytopathic (CPE) appeared++, the supernatant was discarded and half of the The maintenance solution of calf serum is continued to be cultured. When the cytopathic (CPE) appears +++ or more, the cells and supernatant are collected and stored in the refrigerator.

Embodiment 3

[0030] Example 3: Preparation of antigens containing adenovirus fibrin, penton protein and hexon protein

[0031] According to conventional cell culture, after Vero cells have grown into slices, add ADV (1×10 7 TCID50 / ml) was inoculated on Vero cells that had grown into a monolayer, incubated at 33°C, and observed the cytopathic condition every day. When the cytopathic (CPE) appeared++, discard the supernatant and add half of the calf-free Serum maintenance solution continues to be cultured. When the cytopathic (CPE) appears +++ or more, collect the cells and supernatant and store them in the refrigerator.

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Abstract

The invention discloses a preparation method for the immunoglobulin of an adenoviruses anti-Fi, anti-Pb and anti-Hx, which firstly extracts a fibrin, a pentamer protein and a hexon from the adenoviruses as antigens, then purifies the antigens with discontinuous density centrifugation with the virus antigens prepared through the tissue culture, and then uses the prepared purified antigens for establishing and screening the method for the immunoglobulin of high titer adenoviruses anti-Fi, anti-Pb and anti-Hx and the virus neutralization experiment in vitro, etc. while at last the immunoglobulin from the immunoglobulin blood source of high titer adenoviruses anti-Fi, anti-Pb and anti-Hx by means of low temperature ethanol technique, ion exchange chromatography or affinity chromatography is extracted.

Description

1. Technical Field [0001] The present invention belongs to the field of virology and immunology, and relates to a method for preparing medicines, in particular to a method for preparing adenovirus antibodies for prevention and treatment. More specifically, the present invention relates to the source of antigenic substances capable of producing adenovirus-specific immunoglobulins by immunizing animals or humans, the screening method of adenovirus-specific immune plasma or body fluids, and the separation of adenovirus-specific immunoglobulins Preparation methods such as purification. 2. Background technology [0002] Adenovirus (Adenovirus) is a group of DNA viruses that are widely distributed. Can cause human respiratory tract, gastrointestinal tract, urinary system and eye diseases. A few have carcinogenic effects on animals. Adenoviruses are divided into two genera that infect birds and mammals. Human adenoviruses are divided into groups A to G7 according to their physical, chem...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/08
Inventor 余军
Owner TONROL BIOLOGICAL PHARM CO LTD
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