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Method for detection and evaluation of anti-virus-infection activity

An anti-viral and active technology, applied in the field of antibody engineering and anti-viral medicine, can solve the problems that the application efficiency and value of cell infection models and animal protection models are yet to be seen, there are no cell infection and animal protection models, and the operation process is simple , Good reproducibility of results

Active Publication Date: 2013-12-25
FUDAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the cell infection model and animal protection model have been successfully applied to the research of some viruses, such as HIV and HCV, but due to the limitations of the culture and infection conditions of the extracorporeal cell infection system and the heterogeneity of the animal model, the cell infection model and The practical application efficiency and value of the animal protection model remains to be seen
In addition, there are no corresponding cell infection and animal protection models for some virus infections with high species, tissue and cell specificity, such as HBV infection, as well as some new and sudden infection viruses

Method used

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  • Method for detection and evaluation of anti-virus-infection activity
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  • Method for detection and evaluation of anti-virus-infection activity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1: Anti-infective ability of HBV monoclonal antibody and HBIG

[0047] The antiviral activities of the HBV monoclonal antibody and HBIG were detected respectively by using the detection method established in the present invention. HBIG is the human anti-hepatitis B virus globulin produced by Zhongsheng Group, and the humanized anti-hepatitis B virus monoclonal antibody is provided by the Institute of Viral Disease Prevention and Control, Chinese Center for Disease Control and Prevention.

[0048] The specific detection steps are:

[0049] (1) Take isolated blood samples and separate to obtain serum samples of pathogens;

[0050] (2) After mixing the pathogen serum sample with reagent A and reagent B at a ratio of 1:4.5:4.5, add 0.5% Volume test molecules (divided into HBV monoclonal antibody group and HBIG group);

[0051] (3) Stand at room temperature for 3 hours, then centrifuge at 10,000 rpm for 30 minutes;

[0052] (4) The supernatant of the centrifuged m...

Embodiment 2

[0055] Example 2: The antiviral ability of HBIG and anti-HBV monoclonal antibody is related to the formation and deposition of viral complexes

[0056] figure 2 It is the result of the sedimentation specificity experiment.

[0057] Viral DNA was detected according to the method of Example 1, except that serum samples were treated with HBV monoclonal antibody and HBIG without sedimentation treatment, and viral DNA was detected.

[0058] The experimental results showed that the HBV monoclonal antibody BC1 and HBIG in the non-sedimentation treatment group could not clear the virus from the serum. The formation and detection of sedimentary virus complexes are necessary conditions for detection.

Embodiment 3

[0059] Embodiment 3 Specificity experiment of human HBV monoclonal antibody BC1

[0060] image 3 It is a specific experiment for human HBV monoclonal antibody BC1.

[0061] Viral DNA was detected according to the method in Example 1.

[0062] The results showed that the ability of HBV monoclonal antibody to neutralize the virus was 93% (P=0.0001), which was not significantly different from the ability of HBIG to neutralize the virus. The virus clearance ability of HBIG and anti-HBV monoclonal antibody BC1 was specific compared with human serum albumin control.

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Abstract

The invention relates to the antibody engineering and anti-virus medicine fields, and relates to a method for rapid and effective determination of the anti-virus function of recombined human anti-hepatitis B virus antibody. According to the method, settleable virus compound technology is used for establishing a new method for detection and evaluation of anti-virus capability of a molecule. According to the method, the anti-virus and anti-infection capability of the molecule is detected by detection of settleable pathogen compound generation capability of the molecule to be detected, wherein the settleable pathogene compound generation capability of the molecule to be detected is induced in blood serum, and pathogen is virus. The method is used for rapid determination of virus removal and virus infection control capability of an antibody and other anti-infection medicines. The method has the advantages of simple operation process and good result repeatability and is a supplement to the prior art detection and evaluation methods of functions of the anti-virus medicines, and the establishment of the method provides a new choice for research and development of medicines currently having no effective cell and animal infection model virus. The invention also relates to a detection reagent of anti-virus-infection activity, the settleable pathogene compound generated in a detection process and the application of the detection method.

Description

technical field [0001] The invention relates to the fields of antibody engineering and antiviral medicine, in particular to a method for rapidly and effectively measuring the antiviral function of recombinant human anti-hepatitis B virus antibody. Background technique [0002] Human hepatitis B virus (HBV) infection is a global public health problem. According to statistics from the World Health Organization (WHO), more than 2 billion people worldwide have been infected with HBV. Among them, 350 million are suffering from chronic HBV infection (WHO, 2000). my country is a high-incidence area of ​​hepatitis B. The 2006 national viral hepatitis serum epidemiological survey data showed that hepatitis B surface antigen (HBsAg) The positive rate of HBV is 7.18%. Based on this, it is estimated that 93 million people in my country are HBV carriers, of which 30 million are patients with chronic hepatitis B. Studies have also shown that chronic HBV infection is associated with the inci...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68
Inventor 陈力闻玉梅王蕾谢幼华梁米芳
Owner FUDAN UNIV
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