Fraxinus pennsylvanica pollen allergen extract and immersion liquid of fraxinus pennsylvanica pollen allergen extract and preparation method thereof

A technology for allergens and extracts, which is applied in the preparation methods of peptides, allergen antigen components, chemical instruments and methods, etc., can solve the problems of uncertain components, decreased positive rate, poor stability, etc., and achieves short cycle times. , quick effect, high specificity effect

Active Publication Date: 2019-08-02
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

[0006] At present, the allergens used for the allergens of white wax pollen as subcutaneous injection immune preparations have poor stability and uncertain components. When the

Method used

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  • Fraxinus pennsylvanica pollen allergen extract and immersion liquid of fraxinus pennsylvanica pollen allergen extract and preparation method thereof
  • Fraxinus pennsylvanica pollen allergen extract and immersion liquid of fraxinus pennsylvanica pollen allergen extract and preparation method thereof
  • Fraxinus pennsylvanica pollen allergen extract and immersion liquid of fraxinus pennsylvanica pollen allergen extract and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] The identification of embodiment 1 white wax pollen

[0068] 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 and microscopic examination to perform double identification and quality control on the raw material of white wax.

[0069] 1. DNA extraction from white wax pollen

[0070] Tiangen Rapid DNA Extraction and Amplification Kit (Tiangen Biochemical KG203) was used.

[0071] Extraction method: Weigh 5mg of wax 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, centrifuge at 12000r / min for 5min Then take the supernatant and put it in a new centrifuge tube as a DNA template for later use.

[0072] 2. Primer design and synthesis

[0073] The primer sequences are as follows:

[0074] Fra ITS2-F (SEQ ID...

Embodiment 2

[0089] Example 2 Degreasing and Extracting Key Process Steps of Important Parameters of Pyrex Pollen

[0090] 1. Degreasing process parameters of white wax pollen degreasing process

[0091](1) Degrease treatment with the white wax pollen (g) and acetone (ml) at a weight-to-volume ratio (w / v) of 1:2. The fat content of samples of different batches (different collection time) of white wax pollen with different degreasing time was tested to determine the optimal degreasing time.

[0092] Haineng SOX500 fat measuring instrument was used to measure the fat content in the pollen of white wax pollen. Through the method of Soxhlet extraction and dry weighing, the change of pollen weight before and after extraction was compared to obtain the corresponding fat content, and the fat content was compared after degreasing. Pollen for quality control.

[0093] see results figure 2 , it can be seen that after the acetone degreasing time is 30min, the fat content hardly decreases with the...

Embodiment 3

[0104] Embodiment 3 white wax pollen raw material technology

[0105] 1. Pollen collection

[0106] Natural shedding method to collect white wax 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.

[0107] 2. Dry

[0108] 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.

[0109] 3. Degreasing

[0110] 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.

[0111] 4. Dry again

[01...

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Abstract

The invention provides a fraxinus pennsylvanica pollen allergen extract and immersion liquid of the fraxinus pennsylvanica pollen allergen extract and a preparation method thereof. The fraxinus pennsylvanica pollen allergen extract contains an amino acid sequence, which is as shown in SEQ ID NO.4, of fraxinus pennsylvanica pollen allergen protein. The immersion liquid is prepared through technology methods of fraxinus pennsylvanica pollen collecting, drying, degreasing, extracting, ultrafiltration concentrating, freeze-drying, redissolving and the like, and main components of the immersion liquid are a fraxinus pennsylvanica pollen allergen, glycerin, sodium chloride, phenol and the like. The fraxinus pennsylvanica pollen allergen immersion liquid has the character of high specificity, a fraxinus pennsylvanica allergic protein component is fully extracted, a total biological value is stable, a validity period is long, and an aseptic effect is good; and stoste of the immersion liquid ofthe fraxinus pennsylvanica pollen allergen extract can be effectively used for skin prick test diagnosis of allergic reaction diseases, and can be used for in-vitro basophilic granulocyte activationtest diagnosis and specific immunotherapy after being properly diluted so that the allergic reaction diseases induced by fraxinus pennsylvanica pollen can be effectively diagnosed, and can be subjected to specific immunotherapy.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an allergen infusion of white wax pollen, the 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 allergic to grass polle...

Claims

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Application Information

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IPC IPC(8): C07K14/415C07K1/14A61K39/36A61K9/19A61K49/00A61P37/08A61P11/06A61P11/02A61P17/00
CPCC07K14/415A61K39/36A61K9/19A61K49/0006A61P37/08A61P11/06A61P11/02A61P17/00
Inventor 周俊雄尹佳马武强王良录关凯王瑞琦高亚娜
Owner PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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