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Artemisia plant pollen allergen combination, application and kit

A technology for pollen allergies and Artemisia plants, applied in the biological field, can solve the problems of insufficient analysis of allergic asthma and insufficient analysis of allergic asthma

Active Publication Date: 2018-12-21
杭州艾乐吉生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] At present, there is no good method to analyze the risk of allergic asthma, and Artemisia pollen extract is used to detect the level of IgE antibody is not enough to analyze the risk of allergic asthma

Method used

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  • Artemisia plant pollen allergen combination, application and kit
  • Artemisia plant pollen allergen combination, application and kit
  • Artemisia plant pollen allergen combination, application and kit

Examples

Experimental program
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Effect test

Embodiment 1

[0037] Art ar 2 separation and purification, the application of a mouse monoclonal antibody C9-C1 immunoaffinity separation.

[0038] The source of the monoclonal antibody was to immunize 5 mice with 100 μg Artemisia mugwort pollen protein extract (SCXK(HU) 2012-002, China), and inject 50 μg Artemisia protein extract every 7-14 days for a total of six times. According to Western blot and ELISA screening, 3 monoclonal antibodies were obtained, 2 of which bound to Art v 3, and one bound to a protein with a molecular weight of about 16kDa. Take an appropriate amount of CNBr-activated Sepharose 4 Fast Flow (GE healthcare) and add 4 times the volume of cold 1mM HCl, stir and remove the liquid, ddH 2 After washing, mix well with C9-C1 monoclonal antibody dialyzed into coupling buffer, couple overnight at 4°C, measure the concentration of uncoupled protein in the solution to determine the coupling efficiency, and wash with at least 5 times the volume of coupling buffer To remove exc...

Embodiment 2

[0040] The detection of Art v 1 and Art v 3 directly refers to the ImmnuoCAP product manual.

[0041] ImmunoCAP detection of natural allergens Art ar 2 and Art an 7: After the purified natural allergens Art ar2 and Art an 7 were linked with biotin, they were added to streptavidin-conjugated ImmunoCAP (O212) at 37°C at an amount of 0.5 μg / CAP. After incubation for 1 hour, the residual liquid was sucked off for later use (Erwin et al., 2005).

[0042] The specific IgE antibody content was determined by ImmunoCAP (Thermo Fisher, USA), and the detected Artemisia argyi allergen was the extract of Artemisia argyi (w6) and allergen components nArt v 1 (w231), nArt v 3 (w233), nArt ar 2 And nArt an 7, the amount of serum is 40μl, sIgE ≥ 0.35kUA / L is considered positive. The operation process and method, standard curve and positive quality control refer to the ImmnuoCAP 100 product manual. See the test process image 3 . Case information and IgE in vitro test results are shown in T...

Embodiment 3

[0052] At present, there are 7 allergen components of Artemisia pollen (Art v 1–Art v 7), and there are only 2 commercial components for ImmunoCAP detection by ThermoFisher, Sweden: Art v1 and Art v 2. The sum of the measured values ​​of the four allergen components measured by the present invention has a very high correlation (r=0.92, see Figure 4 ). Only a small number of sera lacked the corresponding components, but most of these cases had mild allergic symptoms. These four components are therefore fully representative.

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PUM

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Abstract

The invention discloses an artemisia plant pollen allergen combination for predicting an occurrence risk of asthma of an artemisia allergy patient, an application and a kit. The artemisia plant pollenallergen combination comprises Art v 1, Art ar 2, Art v 3 and Art an 7. The research on 240 artemisia pollen allergy cases discovers that when at least three kinds of allergen specific IgE in the Artv 1, the Art ar 2, the Art v 3 and the Art an 7 are positive in an artemisia pollen allergy case detection result, the occurrence rate of allergic asthma is 57%; when the positive number of the fourallergens is less than 3, the incidence rate of the allergic asthma is 24%; and the former is 2.4 times of the latter. By detecting the allergic patient, the patient with the high occurrence risk of the allergic asthma judged by detection can be possibly cured only by drug control and desensitization treatment, so that the occurrence of the allergic asthma is avoided.

Description

technical field [0001] The invention relates to the technical field related to biotechnology, in particular to an Artemisia plant pollen allergen combination, application and kit for predicting the risk of asthma in Artemisia allergic patients. Background technique [0002] Type 1 allergy is also called type 1 hypersensitivity disease, related diseases include atopic disease, allergic rhinitis, allergic conjunctivitis, atopic dermatitis and allergic asthma (exogenous) and some urticaria , gastrointestinal food reactions and systemic allergic reactions. Type I allergy is widespread worldwide, and patients with atopic disease develop hypersensitivity reactions mediated by IgE antibodies to inhaled or ingested substances (allergens) that are sensitive to non-atopic People are harmless. In addition to atopic dermatitis, usually IgE antibodies mediate hypersensitivity reactions. [0003] The substance that triggers type I allergy is called allergen, which is protein, glycoprot...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/6893G01N2333/47G01N2800/122
Inventor 高中山付婉艺高毕远
Owner 杭州艾乐吉生物科技有限公司
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