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Protein chip for diagnosis allergy and detection method for allergen and antibody

A protein chip and allergen technology, applied in the field of protein chips and detection of allergens that produce antibodies, can solve problems such as inability to distinguish and find the cause of allergic reactions, and achieve the effect of reducing volume, saving time and energy

Inactive Publication Date: 2004-09-08
FOOD BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, for allergic reactions, it can be induced by the crude protein included in the allergen, so it is not necessary to detect a single protein for the detection of allergens
[0009] The conventional approach is to detect anaphylaxis induced by a single allergen, so there is no way to find the cause of all anaphylaxis
Moreover, conventional methods cannot be used to distinguish a specific antibody against a specific allergen from a large number of antibodies

Method used

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  • Protein chip for diagnosis allergy and detection method for allergen and antibody
  • Protein chip for diagnosis allergy and detection method for allergen and antibody
  • Protein chip for diagnosis allergy and detection method for allergen and antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Preparation of protein chip

[0034] 1) Separation of crude protein from allergy-inducing foods

[0035]In order to separate crude protein from allergy-inducing foods, prepare 150 g each of the following substances: milk, egg white, egg yolk, soybean, fresh garlic, garlic treated with artificial gastric acid, uncrushed rice, and barley. These foods are freeze-dried for more than 30 hours to remove water and then rolled into powder. Hexane was added to dry samples at a ratio of 1:5 (w / v), stirred for 30 minutes, and allowed to stand for 30 minutes. The upper layer was decanted and the lower layer was dried. The hexane treatment process was repeated 2 times to remove liquid.

[0036] The liquid-free powder was dissolved in 0.5M NaCl solution at a ratio of 1:4 (w / v), eluted at 4°C for 3 hours, and centrifuged at 15,000 rpm for 1 hour at 4°C. The supernatant was dialyzed against distilled water for 24 hours, lyophilized, and prepared as a protein dissolved i...

Embodiment 2

[0045] Example 2 Using the protein chip of the present invention to detect food allergens

[0046] In order to detect food allergens using the protein chip of the present invention, the protein chip of the present invention was reacted with TBS-T (tris buffer-0.1% Tween 20) and 1% BSA to prevent non-specific reactions. After washing in TBS-T solution for 15 minutes (3 times), the protein chip was reacted with blood sample (60 μl) for 1 hour. After washing, it was reacted with biotin-conjugated anti-human IgE (diluted 1:1000, Vector Co.) at 37°C for 1 hour. To read the protein chip, Cy3 signal was read using ArrayWoRx (Applied Precision Co.), and the program used Imagene (Biodiscovery Co.). The results are shown in Table 2.

[0047] food

[0048] As shown in Table 2, antibodies to egg white, egg yolk, soybean, fresh garlic, and coix were detected in the patient's blood.

Embodiment 3

[0060] Example 3 Using the protein chip of the present invention to detect the type of antibody to a specific food

[0061] In order to detect the type of antibody of a specific food using the protein chip of the present invention, a second antibody containing a different fluorescent type is used for different antibodies. Cy3 and FITC were used as fluorescers for different types of secondary antibodies of the present invention. Fluorescence intensities were measured at wavelengths of 543 nm and 488 nm, respectively.

[0062] Since the results are measured at different wavelengths depending on the antibody, the type of antibody can be determined simultaneously.

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Abstract

This invention is related to the protein chip for allergy diagnosis and the detection method for allergen antibodies. The protein chip of this invention is a solid chip with one crude protein attached to each spot, isolated from various kinds of allergen. By reacting those plates with the sample of allergic patients, the various allergen can be detected at the same time, and the kinds of antibodies related with the allergic reactions caused by the certain allergens.

Description

technical field [0001] The present invention relates to a protein chip for analyzing allergies and a method for detecting allergens that produce antibodies. Background technique [0002] The body's immune system produces antibodies to eliminate antigens, such as foreign substances that enter the body from the outside world. Allergies are the result of a hypersensitive immune response in which antibodies are produced against antigens in a foreign substance and are harmless to most people. As a reaction to the antigen entering the human body, many antibodies are produced, such as IgE that induces allergy, IgG4 that suppresses allergy, and IgG that induces a general immune response. As a result of the interaction between these antibodies, constriction of the bronchi and dilation of the capillaries, stimulation of the secretory glands, and subsequent allergic phenomena such as coughing and runny nose occur. Disorders caused by allergies include bronchial asthma, allergic rhini...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N33/02G01N33/543G01N33/566G01N33/68
CPCG01N33/6854G01N33/53
Inventor 卢建雄
Owner FOOD BIOTECH
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