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Process for preparing allergoid sublingual buccal vaccine using chitosan as adjuvant

An allergen-like, sublingual technology, applied in the field of new vaccine inventions, can solve problems related to immune tolerance, achieve controlled-release gastrointestinal enzyme degradation, simple production process, and reduce hypersensitivity reactions.

Inactive Publication Date: 2006-02-15
SHENZHEN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The study of its therapeutic mechanism is still in its infancy and may be related to immune tolerance

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Example 1: Using biochemical extraction technology to finely extract dust mite allergens

[0014] ①. Sample preparation: Dust mite comes from the Key Laboratory of Microbiology and Genetic Engineering, School of Life Sciences, Shenzhen University. Take 1 gram of Dust mite sample and freeze it in a -70°C refrigerator for 10 minutes. Degrease with cold ether-acetone (1:2). Acetone degreases continuously for 48 hours, during which time the liquid is changed 5 times (every 8-10 hours), and placed under a fume hood to let it dry naturally. All the above operations were carried out on ice bath.

[0015] After degreasing, the samples were weighed and soaked in bicarbonate-saline extract (Coca’s) at a certain ratio. Add the extract at the ratio of 1g sample: 20ml Coca's solution (w / v). At the same time, the samples were fully ground and crushed with a plunger homogenizer. The crushed samples were extracted with Coca's solution for 48hr. During this period, the samples were...

Embodiment 2

[0056] Example 2: Preparation of dust mite allergens

[0057] At 4°C, 1% formaldehyde aqueous solution and 10% D. farinae allergen solution were mixed in equal volumes and stirred gently. Prepare dust mite allergen solution.

Embodiment 3

[0058] Example 3: Identification of dust mite allergens

[0059] Identify the molecular weight of the allergen by SDS-PAGE; detect the IgE reactivity (allergenicity) by competitive ELISA: coat the microwell plate with the allergen solution of D. (PBS; 100mmol / L, pH7.4) to wash the plate. Wells were blocked overnight with 200 [mu]l of PBS containing 10 g / L BSA (BSA / PBS). The five-point calibration curve was operated after serial dilution of the D. farinae allergen solution with BSA / PBS. Immunoglobulin (D. farinae specific IgE) and competitor (diluted calibrator or D. farinae allergen) diluted 1:1 in BSA / PBS were added to the tubes. Take 100 μl from each tube and add it to a microwell plate for 2 hours at 37°C, wash the plate 3 times, add 100 μl horseradish peroxidase-conjugated goat anti-mouse IgE BSA / PBS solution, wash the plate three times at 37°C for 2 hours, wash the plate with 3, 3, 5, 5, and tetramethylbenzidine were used as substrates to determine the binding peroxida...

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PUM

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Abstract

The invention relates to a process for preparing allergoid sublingual buccal vaccine using chitosan as adjuvant, which comprises obtaining natural allergen solution through extraction and purification with biological methods, reacting with formaldehyde, thus obtaining allergoid, detecting biological property through SDS-PAGE and IgE reactivity with competitive ELISA method, viewing antibody titer, finally mixing chitosan with allergoid to thus obtaining chitosan-allergoid vaccine.

Description

technical field [0001] The invention relates to the field of novel vaccine invention, in particular to a preparation method of an allergen-like sublingual oral vaccine with chitosan as an adjuvant. Background technique [0002] Allergic diseases (such as allergic asthma, allergic rhinitis, etc.) are clinically common and frequently-occurring diseases. According to the 2000 WHO / IAACI report, the incidence and mortality of asthma are on the rise in recent years. According to the epidemiological statistics of asthma among adolescents in some cities in the country, the incidence rate of asthma is 3.3%-5.1%, far higher than 1% 10 years ago; at least 10 million children in my country suffer from asthma. [0003] Allergic diseases are caused by various contact or inhalation allergens, most of which are proteins, polypeptides, sugars, and synthetic substances such as drugs. Dust is a strong allergen, which can cause systemic allergic reactions including allergic asthma, allergic de...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/39A61P37/08A61P11/06
Inventor 刘志刚冉丕鑫钟南山喻海琼吉坤美
Owner SHENZHEN UNIV