ELISA kit for capturing food allergen based on IgG3 antibody

A technology of food allergy and kit, applied in the direction of biological testing, material inspection products, measuring devices, etc., can solve the problems of high detection cost, difficult analysis, limiting the development of food allergen detection, etc., to achieve strong specificity and high sensitivity Effect

Inactive Publication Date: 2018-11-20
何韶衡
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  • 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 food allergy detection 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 limits the detection of food allergens. development of

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  • ELISA kit for capturing food allergen based on IgG3 antibody
  • ELISA kit for capturing food allergen based on IgG3 antibody
  • ELISA kit for capturing food allergen based on IgG3 antibody

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preparation example Construction

[0048] In the present invention, the preparation method of the detection plate coated with anti-IgG3 antibody preferably comprises the following steps:

[0049] 1) Diluting the anti-human IgG3 antibody with a coating buffer to obtain a coating solution;

[0050] 2) Add the coating solution obtained in the step 1) to the reaction wells of the detection plate, overnight at 4°C;

[0051] 3) The next day, after washing the detection plate wells overnight in step 2) once, add blocking solution, incubate and wash again.

[0052] In the present invention, the coating buffer is used to dilute the anti-human IgG3 antibody 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. In the present invention, the concentration of the carbonate buffer solution is preferably 0.01-0.2 mol / L. The...

Embodiment 1

[0086] Coating: Dilute the specific anti-human IgG3 monoclonal antibody with carbonate buffer to a coating solution with a protein content of 1.0 μg / ml. Add 0.05ml of coating solution to each reaction well of the polystyrene plate, 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 1.5min each time (referred to as washing, the same below);

[0087]Blocking: Add 0.05ml of PBS buffer solution containing 2% bovine serum albumin to each reaction well, pH: 7.2 (blocking solution), incubate at 15-35°C for 120min, discard the solution in the well, and wash as above;

[0088] Add serially diluted human IgG3 protein (standard wells, see Table 1 for concentrations), PBS buffer (blank control wells), non-allergic volunteer serum (negative control wells) and samples to be tested, i.e. patients with suspected egg, milk and wheat allergies Serum (sample wells, serum were diluted 100 times with PBS buf...

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Abstract

The invention provides an ELISA kit for capturing a food allergen based on an IgG3 antibody, and the ELISA kit belongs to the technical field of food allergen detection. The ELISA kit includes the following reagents: a detection plate coated with the IgG3 antibody, a washing buffer solution, a sample diluent, a standard substance or a positive quality control substance, a biotin-labelled food allergen preparation and a biotin-labelled anti-human IgG3 preparation, an enzyme-labeled avidin solution, a substrate coloring solution, and a stop solution. The ELISA kit of the invention selects the IgG3 to detect the food allergen, and is the prepared ELISA kit based on a capture method; and the ELISA kit has the characteristics of strong sensitivity and good reproducibility, and can process largequantities of samples at one time. Embodiments of the invention prove that the detection sensitivity of the ELISA kit of the invention reaches 1 ng / ml.

Description

technical field [0001] The invention relates to the technical field of allergen detection kits, in particular to a kit for capturing food allergens based on IgG3 antibodies. 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, soybeans, wheat, tree nuts, fish and crustaceans are...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/6893
Inventor 何韶衡何萍
Owner 何韶衡
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