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ELISA kit for capturing airborne allergens based on IgG3 antibody

A kit and allergen technology, which is applied in the field of kits based on IgG3 antibodies to capture airborne allergens, can solve problems such as retention and inability to accurately determine allergic diseases in a timely manner, and achieve high sensitivity and strong detection specificity

Inactive Publication Date: 2018-10-12
THE FIRST AFFILIATED HOSPITAL OF JINZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It can be seen that the current research on airborne allergens in the existing technology stays at the allergic reaction mediated by IgE, which will lead to the inability to timely and accurately determine the occurrence of allergic diseases

Method used

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  • ELISA kit for capturing airborne allergens based on IgG3 antibody
  • ELISA kit for capturing airborne allergens based on IgG3 antibody
  • ELISA kit for capturing airborne allergens based on IgG3 antibody

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preparation example Construction

[0047] In the present invention, the preparation method of the detection plate coated with anti-IgG3 antibody preferably comprises the following steps:

[0048] 1) Diluting the anti-human IgG3 antibody with a coating buffer to obtain a coating solution;

[0049] 2) Add the coating solution obtained in the step 1) to the reaction wells of the detection plate, overnight at 4°C;

[0050] 3) The next day, after washing the detection plate wells overnight in step 2) once, add blocking solution, incubate and wash again.

[0051] 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. The concentration of the carbonate buffer solution is preferably 0.01-0.2 mol / L. The pH value of the carbonate...

Embodiment 1

[0085] 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);

[0086] 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;

[0087]Add gradiently 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. suspected house dust mite and wormwood pollen and serum of patients with Humulus pollen allergy (sample wells, ser...

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Abstract

The invention provides an ELISA kit for capturing airborne allergens based on an IgG3 antibody, and belongs to the technical field of detection of the airborne allergens. The ELISA kit includes the following reagents: a test plate coated with an anti-human IgG3 antibody; a washing buffer; a sample diluent; a standard or a positive quality control; a biotin-labeled airborne allergen preparation anda biotin-labeled anti-human IgG3 antibody preparation; an enzyme-labeled avidin solution; a substrate color developing solution; and a stop solution. The ELISA kit adopts the IgG3 to detect the airborne allergens, and the ELISA kit is an ELISA kit prepared based on a capture method, has the characteristics of high sensitivity and good reproducibility, and can process large quantities of samples at one time. The embodiment proves that the ELISA kit for capturing the airborne allergens based on the IgG3 antibody has a detection sensitivity of 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 airborne 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] Airborne allergens such as pollen particles, fungal spores, dust mites, grain flour, animal ...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/577
CPCG01N33/577G01N33/6854G01N2333/47
Inventor 何韶衡何萍
Owner THE FIRST AFFILIATED HOSPITAL OF JINZHOU MEDICAL UNIV
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