Preparation method and application of dermatophagoides pteronyssinus allergen specific IgG Fab antibody fragment

A house dust mite allergen and antibody fragment technology, applied in the biological field, can solve the problems of dangerous systemic adverse reactions and long treatment time, and achieve the effects of reducing adverse reactions, reducing activation, and controlling inflammatory reactions

Active Publication Date: 2021-11-23
阳江市人民医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

On the other hand, AIT also has limitations: it is suitable for patients with well-controlled disease; the treatment time is long (3-5 years); there is a risk of systemic adverse reactions; sIgG4 drops by nearly 80% after 2 years of stopping treatment

Method used

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  • Preparation method and application of dermatophagoides pteronyssinus allergen specific IgG Fab antibody fragment
  • Preparation method and application of dermatophagoides pteronyssinus allergen specific IgG Fab antibody fragment
  • Preparation method and application of dermatophagoides pteronyssinus allergen specific IgG Fab antibody fragment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1. A method for preparing a house dust mite allergen-specific IgG Fab antibody fragment

[0037] 1. Preparation of candidate serum

[0038] The clinical indications were that after house dust mite-specific immunotherapy was performed on patients allergic to house dust mite (results are shown in Table 1), peripheral blood was aseptically collected according to conventional methods, collected into sterile blood collection tubes, and stored at room temperature (20°C to 25°C). ℃) to stand still for 4 hours, after the serum is separated out, centrifuge at 1500g for 20 minutes, absorb the supernatant serum, and aliquot it into 1.5 ml serum collection tubes to obtain candidate serum.

[0039] Table 1 Conditions of patients with house dust mite allergen-specific immunotherapy

[0040]

[0041] 2. Purification of serum total IgG antibodies from patients with house dust mite allergy after allergen-specific immunotherapy

[0042] (1) Experimental materials:

[0043...

Embodiment 2

[0072] Example 2. A method for preparing house dust mite allergen-specific IgG Fab antibody fragments

[0073] Similar to Example 1, the only difference is that in Step 4, the amount of house dust mite-specific IgG antibody added is 0.9 mL, the amount of agarose-linked papain added is 0.05 mL, and the other parameters are the same as in Example 1.

Embodiment 3

[0074] Example 3. A method for preparing a house dust mite allergen-specific IgG Fab antibody fragment

[0075] Similar to Example 1, the only difference is that in Step 4, the amount of house dust mite-specific IgG antibody added is 0.9 mL, the amount of agarose-linked papain added is 0.2 mL, and the other parameters are the same as in Example 1.

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Abstract

The invention relates to the technical field of biology, and particularly discloses a preparation method and application of a dermatophagoides pteronyssinus allergen specific IgG Fab antibody fragment. The method comprises the following steps: 1) collecting and diluting serum, and filtering to obtain a serum sample; loading the sample on a protein A affinity chromatography column, and then eluting a serum IgG antibody to obtain a serum total IgG antibody; 2) adding dermatophagoides pteronyssinus to a affinity chromatographic column for coupling, adding the serum total IgG antibody in the step 1), and eluting the dermatophagoides pteronyssinus specific IgG antibody; and 3) adding agarose-linked papain into the house dust mite specific IgG antibody, uniformly mixing, incubating, centrifuging, and taking supernatant, thereby obtaining the house dust mite specific immunoglobulin G. The dermatophagoides pteronyssinus allergen specific IgG Fab antibody fragment can compete with IgE to bind to dermatophagoides pteronyssinus and reduce the formation of a dermatophagoides pteronyssinus-IgE compound, so that the binding of the dermatophagoides pteronyssinus-IgE compound to effector cells expressing IgE low-affinity receptors and IgE high-affinity receptors is reduced, the activation of sensitized cells is reduced, and the occurrence of inflammatory response is controlled.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a preparation method and application of a house dust mite allergen-specific IgG Fab antibody fragment. Background technique [0002] Asthma is a chronic airway inflammation involving mast cells, eosinophils, basophils and T lymphocytes. Epidemiological surveys show that the global prevalence of asthma is as high as 5% to 16%. More than 50% of adult asthma patients and 80% of children with asthma are sensitive to allergens, and dust mite is the main allergen of allergic asthma patients in my country, and 87.2% of allergic patients are allergic to more than one dust mite , and the degree of sensitization was positively correlated with the severity of asthma. [0003] The prevention and treatment of allergic asthma are closely related: on the one hand, identify the allergen as much as possible and avoid contact with it; According to the treatment guidelines, the condition can be contr...

Claims

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Application Information

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IPC IPC(8): C07K16/18C07K16/06C07K1/22C12P21/06A61K39/395A61P37/08A61P11/06A61P11/02A61P17/00
CPCC07K16/18C07K16/065C12P21/06A61P37/08A61P11/06A61P11/02A61P17/00C07K2317/55C07K2317/52C07K2317/56C07K2317/76Y02A50/30
Inventor 冯木林
Owner 阳江市人民医院
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