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Ragweed pollen allergen extract, ragweed pollen allergen extraction liquid and preparation method of extract

A technology of allergens and extracts, which is applied in the direction of allergen antigen components, pharmaceutical formulations, powder delivery, etc., can solve the problems of protein content and biological potency decrease, positive rate decrease, poor stability, etc., to avoid detection Effects of false positives, improved stability, and short cycles

Active Publication Date: 2019-06-28
PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the stability of ragweed pollen allergens used as subcutaneous injection immune preparations is poor, and the protein content and biological potency will decrease with time. The reagents with uncertain components are used to diagnose allergic diseases induced by ragweed pollen after a certain period of storage When , the positive rate decreases and the accuracy is low

Method used

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  • Ragweed pollen allergen extract, ragweed pollen allergen extraction liquid and preparation method of extract
  • Ragweed pollen allergen extract, ragweed pollen allergen extraction liquid and preparation method of extract
  • Ragweed pollen allergen extract, ragweed pollen allergen extraction liquid and preparation method of extract

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2

[0086] Example 2 Degreasing of ragweed pollen and determination of important parameters in key process steps of extraction

[0087] 1. Degreasing process parameters of ragweed pollen

[0088] (1) The ragweed pollen (g) and acetone (ml) were degreased at a weight-to-volume ratio (w / v) of 1:2. The fat content of different batches (different collection time) of ragweed pollen with different defatting time samples was tested to determine the optimal defatting time.

[0089] The fat content of ragweed pollen was measured by the Sea Energy SOX500 fat analyzer, and the weight change of the pollen before and after extraction was compared by Soxhlet extraction and dry weighing method to obtain the corresponding fat content, and the fat content was compared with the fat content after degreasing Pollen for quality control.

[0090] see results figure 2 , it can be seen that after the acetone degreasing time is 30min, the fat content hardly decreases with the prolongation of the degre...

Embodiment 3

[0101] Embodiment 3 ragweed pollen raw material technology

[0102] 1. Pollen collection

[0103] Collect ragweed pollen by natural shedding. Normal temperature or vacuum or fluidized bed drying, until the pollen no longer sticks. Pass the dried pollen through a 150-250 mesh sieve, preferably a 180 mesh sieve in this embodiment.

[0104] 2. Dry

[0105] Spread the pollen in a ventilated and dry place to dry naturally, or vacuum dry, or dry in a fluidized bed for 6-48 hours, until the pollen no longer adheres.

[0106] 3. Degreasing

[0107] The pollen (g) obtained above and acetone (ml) are subjected to degreasing treatment at a weight-to-volume ratio (w / v) of 1:5-1:1. In this embodiment, it is preferably 1:2. Stir or oscillate to degrease for 30 minutes, let stand to separate layers, and observe the clarification of the upper liquid. Pour out the upper layer, add new acetone, and repeat degreasing until the upper layer is clear.

[0108] 4. Dry again

[0109] Spread ...

Embodiment 4

[0116] Embodiment 4 Preparation process of ragweed pollen allergen stock solution (lyophilized product)

[0117] 1, according to embodiment 3 ragweed pollen raw material process preparation ragweed pollen raw material.

[0118] 2. Leaching

[0119] Add the defatted and dried ragweed pollen to 10 L at a weight-to-volume ratio of 1:50 to 1:10, preferably 1:20 in this example, and stir in pH 7.9 to 8.2 phosphate-saline buffer solution at 2 to 8°C for 22 ~26h for leaching. Wherein, the formula of preparing 1000ml phosphate-saline buffer (pH 8.2) as an example is shown in Table 4. Phosphate-saline buffer solution is fully dissolved, sterilized and filtered, placed at 2-8°C, and the storage period does not exceed 30 days.

[0120] Table 4 Phosphate-saline buffer (pH 8.2) formulation

[0121]

[0122] 3. Solid-liquid separation

[0123] Take the extracted extract, add all the extract into the centrifuge bucket, adjust the balance, centrifugal force 8000-12000g, centrifuge tem...

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Abstract

The invention relates to a ragweed pollen allergen extract, ragweed pollen allergen extraction liquid and a preparation method of the extract. The ragweed pollen allergen extraction liquid includes ragweed pollen allergens containing the amino acid sequences shown in SEQIDNO.4 to SEQIDNO.9. The ragweed pollen allergen extraction liquid is prepared through ragweed pollen collection, drying, degreasing, extraction, ultra-filtration concentration, freeze drying, re-dissolving and the like and mainly includes ragweed pollen allergens, glycerinum, sodium chloride and the like. The ragweed pollen allergen extraction liquid has the advantage of being high in specificity, allergenic protein components of ragweed are fully extracted, the total biological value is stable, the service life is long, and the sterile effect is good. The stoste of the ragweed pollen allergen extraction liquid can be effectively used for skin prick test diagnosis of inspiratory allergosis, in-vitro basophilic granulocyte activation test diagnosis, intracutaneous test diagnosis and specific immunotherapy after appropriate dilution, and can effectively diagnose inspiratory allergosis induced by ragweed pollen and conduct specific immunotherapy on the inspiratory allergosis.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a standardized allergen extract of ragweed pollen, its solution and a preparation method thereof. Background technique [0002] People's understanding of allergic diseases began with "hay fever", which is also known as allergic rhinitis. In 1911, Noon and Freeman used pollen extract to treat hay fever for the first time, and the history of allergenic disease treatment began from then on. At present, allergic diseases are one of the major global health problems. More than 25% of the population in industrialized countries is troubled by allergic asthma, allergic rhinoconjunctivitis and allergic dermatitis, among which allergic asthma is the most common. Inhalation of allergenic pollen is the most important factor in triggering allergic asthma and other respiratory allergic diseases. Surveys such as VRTA-L.A found that nearly 50% of patients with allergic diseases were allergic to gr...

Claims

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Application Information

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IPC IPC(8): A61K39/36A61K49/00A61K9/19A61K9/08A61K47/10A61P37/08A61P11/06A61P11/02A61P17/00
Inventor 周俊雄尹佳高亚娜王茜文利平路蒨
Owner PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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