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Artemisia sieversiana pollen allergen extract and immersion liquid and preparation method thereof

A technology of Artemisia annua and allergen, applied in the biological field, can solve the problems of decreased protein content and biological titer, decreased positive rate, low accuracy, etc., and achieves improved stability and sustained release effect, short cycle and effective results. quick effect

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 allergens used for subcutaneous injection of Artemisia grandis pollen have poor stability, and the protein content and biological potency will decrease with time. The reagents with uncertain components can be used to diagnose metamorphosis induced by Artemisia grandis pollen after being stored for a certain period of time. When reflecting the disease, the positive rate is reduced and the accuracy is low

Method used

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  • Artemisia sieversiana pollen allergen extract and immersion liquid and preparation method thereof
  • Artemisia sieversiana pollen allergen extract and immersion liquid and preparation method thereof
  • Artemisia sieversiana pollen allergen extract and immersion liquid and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0068] The identification of embodiment 1 Artemisia grandis pollen raw material

[0069] Since the identification and purity of raw materials are extremely important for subsequent allergen diagnosis and therapeutic preparations, this example uses DNA-specific sequence detection plus microscopy to perform double identification and quality control of Artemisia grandis raw materials.

[0070] 1. DNA extraction from Artemisia grandis

[0071] Tiangen Rapid DNA Extraction and Amplification Kit (Tiangen Biochemical KG203) was used.

[0072] Extraction method: Weigh 5mg of Artemisia grandis pollen sample and place it in a 1.5mL centrifuge tube, add 100μl of Buffer 1, grind the sample thoroughly with a disposable grinding pestle, add 100μl of Buffer 2, shake and mix well, and centrifuge at 12000r / min After 5 minutes, take the supernatant and put it in a new centrifuge tube as a DNA template for later use.

[0073] 2. Primer design and synthesis

[0074] The primer sequences are as...

Embodiment 2

[0092] Example 2 Degreasing and extraction of Artemisia grandis pollen key parameters determination of key process steps

[0093] 1. Degreasing parameters of Artemisia grandis pollen degreasing process step

[0094] (1) Degreasing treatment of Artemisia grandis pollen (g) and acetone (ml) at a weight-to-volume ratio (w / v) of 1:2. The fat content of different batches (different collection time) of Artemisia grandis pollen with different defatting time samples was tested to determine the optimal defatting time.

[0095] Haineng SOX500 fat measuring instrument was used to measure the fat content in the pollen of Artemisia grandis, through Soxhlet extraction and dry weighing method, the change of pollen weight before and after extraction was compared to obtain the corresponding fat content, and the fat content was used according to the fat content after degreasing Post-pollen quality control.

[0096] see results figure 2 , it can be seen that after the acetone degreasing time...

Embodiment 3

[0107] Embodiment 3 artemisia grandis pollen raw material technology

[0108] 1. Pollen collection

[0109] Natural shedding method, collecting Artemisia grandis pollen. 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.

[0110] 2. Dry

[0111] 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.

[0112] 3. Degreasing

[0113] 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.

[0114] 4....

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Abstract

The invention provides an artemisia sieversiana pollen allergen extract and immersion liquid and a preparation method thereof. The artemisia sieversiana pollen allergen extract contains an artemisia sieversiana pollen allergen protein of which the amino acid sequence is shown in SEQ ID NO.4-SEQ ID NO.10. The artemisia sieversiana pollen allergen extract is prepared through artemisia sieversiana pollen collection, drying, degreasing, extraction, ultrafiltration concentration, freeze drying, redissolving and other processes. The artemisia sieversiana pollen allergen extract has the advantages that the specificity is high, the extraction of an artemisia sieversiana allergenic protein component is sufficient, the total biological value is stable, the period of validity is long, and the sterileeffect is good; a stock solution of the artemisia sieversiana pollen allergen extract can be effectively used for skin prickling test diagnosis of allergic diseases, can be effectively used for in-vitro basophilic granulocyte activation test diagnosis and specific immunotherapy after being diluted appropriately, and can effectively diagnose the allergic diseases induced by artemisia sieversiana pollen and carry out specific immunotherapy on the allergic diseases.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a standardized allergen extract from Artemisia grandis pollen, an infusion 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 all...

Claims

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Application Information

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IPC IPC(8): A61K39/36A61K9/19A61P37/08A61K49/00C07K14/415C07K1/34
Inventor 周俊雄尹佳王瑞琦关凯邢旭亚
Owner PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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