Immunoglobulin antibody against SARS-CoV and its preparing method

A technology of immunoglobulin and antibody, which is applied in the direction of antiviral immunoglobulin, antibody, liquid transportation, etc., can solve the problems of restricting application and development, superficial understanding, etc., and achieve the effect of good specificity and sensitivity

Inactive Publication Date: 2007-02-14
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Based on reports from Hong Kong, Singapore and my country, the serum of convalescent patients with SARS has the effect of treating SARS, but there are certain problems in the wide application of serum from convalescent patients with SARS, and people's understanding of SARS-CoV is still very superficial, especially in the There are certain problems in security and other aspects, which restrict its application and development

Method used

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Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0027] Experimental example 1 refined horse anti-SARS-CoV immunoglobulin F (ab') 2 Antibody identification

[0028] 1) Refined horse anti-SARS-CoV immunoglobulin F(ab') 2 Antibodies and purified horse anti-SARS-CoV immunoglobulin antibodies, the antibody titer of convective electrophoresis reaches 1:128 or more, and the indirect ELISA antibody titer reaches 1:12800 or more;

[0029] 2) Refined horse anti-SARS-CoV immunoglobulin F (ab') 2 The titration of the neutralizing titer of the antibody is first determined by titrating SARS-CoV on Vero cells to be 10 6-8 , select 200TCID 50 Detection of horse anti-SARS-CoV plasma and refined horse anti-SARS-CoV immunoglobulin F(ab′) by antibody dilution method 2 Neutralizing titers of antibodies. Or detect horse anti-SARS-CoV plasma, refined horse anti-SARS-CoV immunoglobulin F (ab') by plaque subtraction neutralization assay 2 The neutralizing titer of the antibody is above 1:10000;

experiment example 2

[0030] Experimental example 2 Evaluation of therapeutic and preventive effects of refined horse anti-SARS-CoV immunoglobulin

[0031] 1) To establish the SARS-CoV infection Vero-6 cell model, add 4×10 5 / well Vero-6, add 100CTID after culturing for 24h 50 BJ01 SARS-CoV200ul, add 0.1mg / 100mL horse anti-SARS-CoV immunoglobulin after 24h to treat SARS-CoV-infected cells, observe 48h by cytopathic method, MTT method and plaque subtraction method to detect horse anti-SARS-CoV immunity Globulin has a good effect on treating SARS-CoV;

[0032] 2) Equine anti-SARS-CoV immunoglobulin 0.1mg / 100mL was added to 4×10 5 / well Vero-6 cells / 16-well cell culture plate, use 100CTID for 12h 50 BJ01 SARS-CoV 200ul infected Vero-6 cells, observed 72h by cytopathic method, MTT method and plaque subtraction method to detect the refined horse anti-SARS-CoV immunoglobulin has a good effect on preventing SARS-CoV;

[0033] 3) By 1×10 for each rhesus monkey 8 BJ01 strain SARS-CoV was infected by int...

Embodiment 1

[0036] Example 1 Horse anti-SARS-CoV immunoglobulin F (ab') 2 Antibody preparation

[0037] 1. The cultivation, inactivation and purification of SARS-CoV:

[0038] 1) Recovery and cultivation of green monkey kidney cells (Vero cells), the Vero cell strains were frozen from the cell seed bank, cultured with DMEM medium and 10% calf serum, and Vero cells for cultivating SARS-CoV were provided;

[0039] 2) SARS coronavirus (SARS-CoV) titration, adding identified and frozen SARS-CoV to cultured Vero cells, amplifying and culturing SARS-CoV, using tissue culture to measure the virulence of SARS-CoV virus, and its half infection rate (TCID 50 ) is 10 6 -10 8 , subpackage and freeze the titrated SARS-CoV;

[0040] 3) adding titrated SARS-CoV to Vero cells covered with monolayer, and culturing SARS-CoV with serum-free DMEM to produce SARS-CoV;

[0041] 4) Collect the SARS-CoV culture medium after Vero cells are completely lesioned, freeze and thaw three times (frozen at -37°C) o...

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PUM

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Abstract

The present invention relates to anti-SARS-CoV immune globulin F(ab' )2 antibody for treatment, prevention and diagnosis of SARS-CoV. The present invention features that the antibody is horse anti-SARS-CoV immune globulin F(ab' )2 antibody. The present invention also discloses the preparation process of the antibody. The antibody of the present invention can kill SARS-CoV specifically, and has excellent effect of preventing and treating SARS.

Description

technical field [0001] The invention relates to an anti-SARS-CoV immunoglobulin antibody used for treating, preventing and diagnosing SARS-CoV, belonging to the field of biopharmaceuticals. Background technique [0002] After the World Health Organization issued a global warning on atypical pneumonia (atypical pneumonia) on March 12 this year, it was subsequently named severe acute respiratory syndrome (Severe Acute Respiratory Syndrome, SARS). Under the great attention, coordination and organization of the WHO, the pathogen causing SARS was quickly identified as a new or mutated coronavirus, and was named SARS-CoV accordingly. Since the outbreak of SARS, it has quickly spread around the world, and has now spread to more than 30 countries and regions in the world, seriously affecting people's health and life, and becoming the number one enemy that endangers mankind most in the early 21st century. Horse antiserum (also known as horse antiimmune globulin) has been an effectiv...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/10A61K39/395A61K9/12A61K9/08A61K9/10A61P31/14A61P11/00
Inventor 王希良杜新安石辛甫张松乐刘建源李岩赵光宇田光周华
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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