ELISA kit for detecting food allergen based on IgG2 antibody

A technology for food allergy and antibody detection, applied in biological testing, material inspection products, etc., can solve the problems of difficult analysis, high detection cost, and slow detection speed, and achieve the effects of strong detection sensitivity, improved sensitivity, and wide application range

Inactive Publication Date: 2018-11-13
何韶衡
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Detect and identify allergens in food by analyzing the characteristics of allergen proteins, but special large-scale precision instruments are required, the detection cost is high, and the analysis is difficult. Special testing personnel and analysts are required.
[0005] To sum up, although there are many methods for the detection of food allergens currently studied, they all have the problems of slow detection speed, high cost, and are not suitable for batch detection, and require specific analytical instruments for machine-reading, which restricts the detection of food allergens. Development of original assays
Most importantly, no food allergen-specific IgG2 detection kit currently exists

Method used

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  • ELISA kit for detecting food allergen based on IgG2 antibody
  • ELISA kit for detecting food allergen based on IgG2 antibody
  • ELISA kit for detecting food allergen based on IgG2 antibody

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Experimental program
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preparation example Construction

[0049] In the present invention, the preparation method of the detection plate coated with food allergen preferably comprises the following steps:

[0050] 1) diluting the food allergen with a coating buffer to obtain a food allergen coating solution;

[0051] 2) Add the coating solution obtained in the step 1) to the reaction wells of the detection plate, overnight at 4°C;

[0052] 3) The next day, after washing the detection plate wells overnight in step 2) once, add blocking solution, incubate and wash again.

[0053] In the invention, the food allergen is diluted with the coating buffer solution to obtain the coating solution. In the present invention, the dilution method is not particularly limited, and a dilution technique known to those skilled in the art can be used. In the present invention, the coating buffer is preferably carbonate buffer. The concentration of the carbonate buffer solution is preferably 0.01-0.2 mol / L. The pH value of the carbonate buffer soluti...

Embodiment 1

[0088]Coating: dilute egg white, wheat and milk food allergens with carbonate buffer solution to a food allergen coating solution with a total protein concentration of 10 μg / ml; dilute human IgG2 protein with carbonate buffer solution to obtain a standard Coating solution (see Table 1 for protein concentration), add 0.05ml of coating solution to each reaction well of the polystyrene plate, and overnight at 4°C. On the next day, discard the solution in the wells, add 0.35ml washing buffer to each well, wash 3 times, and incubate for 30s each time (referred to as washing, the same below);

[0089] Blocking: Add 0.05ml of PBS buffer (blocking solution) containing 2% bovine serum albumin to each reaction well, pH: 7.2, incubate at 15-35°C for 120min, discard the solution in the well, and wash as above;

[0090] Add samples: serum from patients with suspected egg allergy, serum from patients with wheat allergy, serum from patients with milk allergy (serum to be tested), normal huma...

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Abstract

The invention provides an ELISA kit for detecting food allergen based on IgG2 antibody, and belongs to the technical field of food allergen. The kit is prepared from the following reagents: a detection plate coated with food allergen; a wash buffer solution, a sample diluent, a standard or a positive quality control, a biotin labeled anti-human IgG2 antibody preparation, an enzyme labeled avidin solution, a substrate developing solution and a stop solution. According to the ELISA kit provided by the invention, IgG2 is adopted to detect food allergen for the first time; the ELISA kit prepared based on an indirect method has the characteristics of high detection sensitivity, high accuracy and good reproducibility, and can be used for treating a large batch of samples once. Embodiments provethat the detection sensitivity of the kit provided by the invention can reach 0.5ng / ml.

Description

technical field [0001] The invention relates to the technical field of food allergen detection, in particular to a kit for detecting food allergens based on IgG2 antibodies, that is, a food allergen-specific IgG2 detection kit. Background technique [0002] The incidence of allergic diseases accounts for more than 30% of the world's total population, and is listed by the World Health Organization as one of the four major non-infectious diseases in the 21st century. In recent years, the incidence of allergic diseases has increased significantly and has become a public health issue of global concern. The epidemiological survey of allergic diseases by Guo Jia et al. shows that 22% of the total population of 1.2 billion suffer from allergic diseases. For example, allergic rhinitis, asthma, conjunctivitis, eczema, food allergy, drug allergy and severe anaphylaxis, etc., seriously affect people's quality of life and life safety. [0003] Food allergens such as eggs, peanuts, milk...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53
CPCG01N33/53
Inventor 何韶衡何萍
Owner 何韶衡
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