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Kochia scoparia(l.)schrad. pollen allergen extract and extract liquid and preparation method of extract liquid

An allergen and pollen technology, applied in the directions of allergen antigen components, drug combinations, pharmaceutical formulations, etc., can solve the problems of decreased protein content and biological titer, decreased positive rate, poor stability, etc., to improve stability and performance. Sustained release, short cycle, high specificity

Active Publication Date: 2019-09-06
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 subcutaneous injection immunization preparations used for Kochia pollen allergens is poor, and the protein content and biological potency will decrease with time. The reagents with uncertain components can be used to diagnose allergic diseases induced by Kochia pollen after being stored for a certain period of time. When , the positive rate decreases and the accuracy is low

Method used

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  • Kochia scoparia(l.)schrad. pollen allergen extract and extract liquid and preparation method of extract liquid
  • Kochia scoparia(l.)schrad. pollen allergen extract and extract liquid and preparation method of extract liquid
  • Kochia scoparia(l.)schrad. pollen allergen extract and extract liquid and preparation method of extract liquid

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] The identification of embodiment 1 Kochia 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 carry out double identification and quality control of Kochia scoparia raw materials.

[0069] 1. Kochia pollen DNA extraction

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

[0071] Extraction method: Weigh 5mg Kochia pollen sample and place it in a 1.5mL centrifuge tube, add 1100μl of Buffer, grind the sample thoroughly with a disposable grinding pestle, add 100μl of Buffer 2, shake and mix, centrifuge at 12000r / min for 5min Take the supernatant in a new centrifuge tube and use it as a DNA template for later use.

[0072] 2. Primer design and synthesis

[0073] The primer sequences are as follows:

[0074] 1. Primer Design The primer seque...

Embodiment 2

[0095] Example 2 Degreasing of Kochia pollen and Determination of Key Parameters in Key Process Steps of Extraction

[0096] 1. Degreasing Process Step Parameters of Kochia Pollen

[0097] (1) Degreasing the Kochia 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 Kochia pollen samples with different degreasing time was tested to determine the optimal degreasing time.

[0098] The fat content in Kochia pollen was measured with 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.

[0099] 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 degreasing ...

Embodiment 3

[0110] Embodiment 3 Kochia pollen raw material technology

[0111] 1. Pollen collection

[0112] Natural shedding method, collecting Kochia 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.

[0113] 2. Dry

[0114] 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.

[0115] 3. Degreasing

[0116] 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.

[0117] 4. Dry again

[0118] S...

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Abstract

The invention provides an kochia scoparia(l.)schrad. pollen allergen extract and extract liquid and a preparation method of the extract liquid. The kochia scoparia(l.)schrad. pollen allergen extract contains a sensitizing protein Koc S1' containing an amino acid sequence shown in SEQ ID NO.4. The kochia scoparia(l.)schrad. pollen allergen extract liquid contains a therapeutic or diagnostic effective dose of the kochia scoparia(l.)schrad. pollen allergen. The extract liquid can also be prepared by process methods of kochia scoparia(l.)schrad. pollen collection, drying, degreasing, extraction, ultrafiltration concentration, freeze-drying and re-dissolution and the like. The kochia scoparia(l.)schrad. pollen allergen extract liquid has the advantages of high specificity, a kochia scoparia(l.)schrad. sensitizing protein component is extracted adequately, the total biological potency is stable, the period of validity is long, and the sterile effect is good; the kochia scoparia(l.)schrad. pollen allergen extract liquid can be effectively used in skin prick test diagnosis of allergic diseases, in-vitro basophil activation test diagnosis and specific immunotherapy, and can effectively diagnose allergic diseases induced by gkochia scoparia(l.)schrad. pollen and performs specific immunotherapy.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a Kochia pollen allergen extract, its extract 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 pollen. [00...

Claims

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Application Information

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IPC IPC(8): A61K49/00A61K39/36A61P37/08
CPCA61K39/36A61K49/0006A61P37/08
Inventor 周俊雄尹佳王良录王瑞琦顾建青支玉香李宏汤蕊
Owner PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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