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Method for detecting allergen ovomucoid or ovotransferrin in influenza vaccine quantificationally

A technology of ovomucoid and transferrin, which is applied in the direction of measuring devices, biological testing, material inspection products, etc., can solve the problems of no influenza vaccine allergen content, insufficient recognition, etc., and achieve easy quality control and specificity Strong and repeatable effect

Inactive Publication Date: 2012-08-29
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, the new allergens of ovomucoid or ovotransferrin in influenza vaccines have not been fully recognized at home and abroad, and there are no regulations on the content of these two types of allergens in influenza vaccines

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Embodiment 1: Quantitative detection of ovomucoid in influenza virus stock solution:

[0018] Using ovomucoid as immunogen, using conventional hybridoma technology, through cell fusion, screening and cloning, a total of 8 anti-ovomucoid monoclonal antibodies were obtained, which were named FMU-OM1 ~ FMU-OM8.

[0019] After antibody pair screening, the FMU-OM5 monoclonal antibody was used as the coating antibody, and the FMU-OM6 monoclonal antibody labeled with horseradish peroxidase was used as the enzyme-labeled antibody. With pH9.6, 0.05mol / L Na 2 CO 3 -NaHCO 3 Dilute the coated antibody FMU-OM5 to 5 mg / L with buffer solution, add 100 μl / well to a 96-well plate, keep it moist at 4°C for 24 hours. Wash the 96-well plate three times with washing buffer, add buffer containing 1% bovine serum albumin, and block at room temperature for 30 minutes. Wash the plate 3 times, add the sample of influenza virus stock solution extracted from the diluted chicken embryo allantoi...

Embodiment 2

[0020] Embodiment 2: Quantitative detection of ovotransferrin in influenza virus stock solution:

[0021] Using ovotransferrin as an immunogen, 18 strains of monoclonal antibodies against ovotransferrin were obtained by conventional hybridoma technology through cell fusion, screening and cloning, which were named FMU-OT1~FMU-OT18.

[0022] After antibody pair screening, the FMU-OT12 monoclonal antibody was used as the coating antibody, and the FMU-OT6 monoclonal antibody labeled with horseradish peroxidase was used as the enzyme-labeled antibody. With pH9.6, 0.05mol / L Na 2 CO 3 -NaHCO 3 Dilute the coated antibody FMU-OT12 to 5 mg / L with buffer solution, add 100 μl / well to a 96-well plate, keep it moist at 4°C for 24 hours. Wash the 96-well plate three times with washing buffer, add buffer containing 1% bovine serum albumin, and block at room temperature for 30 minutes. Wash the plate 3 times, add the sample of influenza virus stock solution extracted from the diluted chick...

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PUM

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Abstract

The invention discloses a method for detecting allergen ovomucoid or ovotransferrin in influenza vaccine quantificationally. Based on the preparation of anti-ovomucoid or anti-ovotransferrin monoclonal antibody, after paired antibodies are screened, a double antibody sandwich ELIsA (enzyme linked immunosorbent assay) adsorption test is established to quantificationally detect allergen ovomucoid or ovotransferrin in influenza vaccine. The method aims to reduce the incidence rate of allergic reaction of inoculated people caused by ovomucoid or ovotransferrin remaining in influenza vaccine prepared in a chick embryo allantoic cavity in a purified manner, has the advantages of strong specificity, high sensitivity, good repeatability and simplicity and convenience in implementation, and is very suitable for quickly and quantificationally detecting ovomucoid and ovotransferrin in food or influenza vaccine in disease control.

Description

technical field [0001] The invention belongs to the technical field of immune analysis, and relates to a protein detection method, in particular to a method for quantitatively detecting new allergens ovomucoid or ovotransferrin in influenza vaccines by using a double-antibody sandwich enzyme-linked immunosorbent assay. Background technique [0002] Influenza is the disease with the greatest potential for global epidemics. Several world pandemics of influenza in the last century have caused huge losses to the world. Since the beginning of this century, the World Health Organization has continuously released human cases of H5N1 bird flu, especially the H5N1 bird flu that has been released around the world and the new HIN1 flu (swine flu) that broke out in recent years, which may cause a global pandemic. Influenza vaccine is the most effective way to prevent the occurrence and spread of influenza. At present, the main way of producing influenza vaccines in various countries in...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/577G01N33/543
Inventor 张春梅李琦宋朝君金伯泉李永明徐竹蔚陈丽华杨琨张赟方亮易静周幸春马樱刘蓓张宇丝刘志佳
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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