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Nasal cavity excitation reagent based on recombinant dermatophagoides farinae Der f2 protein

A technology of dust mite and protein, applied in the field of biomedicine, to achieve the effect of ensuring safety and accuracy

Pending Publication Date: 2021-10-22
ZHONGSHAN HOSPITAL FUDAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There is currently no standardized nasal challenge reagent for this allergen

Method used

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  • Nasal cavity excitation reagent based on recombinant dermatophagoides farinae Der f2 protein
  • Nasal cavity excitation reagent based on recombinant dermatophagoides farinae Der f2 protein
  • Nasal cavity excitation reagent based on recombinant dermatophagoides farinae Der f2 protein

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Experimental program
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Effect test

Embodiment

[0018] 1. Materials and methods

[0019] 1.1. Materials

[0020] 1.1.1. Sera from patients with dust mite allergy Twenty-six patients with allergic rhinitis underwent skin prick test for common allergens, of which 24 cases had positive prick reaction to dust mite and 2 cases had negative reaction to dust mite. Take 5mL1 venous blood, separate the serum, and store it at -80°C. The Der f2-specific IgE-positive serum panel (panel) came from PlasmaLabInternational and GREER LABORATORIES, INC., USA.

[0021] 1.1.2. Clinical prick test materials: Allergopharma products for pricking liquid and pricking needles for dust mite allergens.

[0022] 1.1.3. BCA protein quantification kit was purchased from Pierce Company.

[0023] 1.1.4. Prokaryotic expression of Dermatophagoides farinae protein and purified wild-type Der f2.

[0024] 1.1.5. ELISA reagents and materials Coating reagent: 0.05M carbonate buffer (pH9.0). Washing solution: 1PBS, add Tween-20 to 0.02% concentration. Blocki...

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Abstract

The invention relates to a nasal cavity excitation reagent based on recombinant dermatophagoides farinae Der f2 protein, and belongs to the technical field of biological medicine. The invention provides an application of a recombinant dermatophagoides farinae Der f2 protein in preparation of an allergen nasal cavity excitation reagent. The invention also discloses an application of the recombinant dermatophagoides farinae Der f2 protein in preparation of a diagnostic reagent for diagnosing allergic rhinitis. The reagent comprises a spray, a drop or an injection; the key allergenic peptide fragment of the recombinant dermatophagoides pteronyssinus protein is provided, and it is proved that the key allergenic peptide fragment can cause degranulation of in-vivo mast cells. The protein is applied to a nasal cavity excitation experiment to check whether corresponding IgE exists in nasal mucosa or not, and clear diagnosis can be achieved. The recombinant protein can be prepared into standard concentration to meet the requirement of concentration gradient, so that the safety and accuracy of nasal cavity excitation test are ensured.

Description

technical field [0001] The invention relates to a nasal stimulation reagent based on recombinant Der f2 protein of dust mite farinae, belonging to the technical field of biomedicine. Background technique [0002] Allergic rhinitis is a common disease with an increasing incidence year by year. A definite diagnosis requires nasal provocation of allergens. Nasal provocation testing (NPT) is currently the internationally recognized "golden test" for diagnosing allergic rhinitis allergens. It is widely used in the evaluation of the effectiveness of therapeutic drugs and the study of the pathophysiological mechanism of nasal stimulation to allergens. Dust mites are common respiratory allergens in southern China. There is currently no standardized nasal challenge reagent for this allergen. Therefore, there is an urgent need in the technical field to obtain a dust mite-like respiratory allergen for the allergen nasal challenge test. Contents of the invention [0003] The purpos...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/00G01N33/68
CPCA61K49/0004G01N33/6893G01N2333/43582G01N2800/14
Inventor 徐枫张利能
Owner ZHONGSHAN HOSPITAL FUDAN UNIV
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