A kind of Kochia pollen allergen extract, its extract and its preparation method

An allergen and pollen technology, applied in the biological field, can solve the problems of decreased protein content and biological titer, decreased positive rate, low accuracy, etc., to achieve the effects of avoiding false positive detection, improving stability, and shortening the cycle

Active Publication Date: 2021-08-03
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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  • A kind of Kochia pollen allergen extract, its extract and its preparation method
  • A kind of Kochia pollen allergen extract, its extract and its preparation method
  • A kind of Kochia pollen allergen extract, its extract and its preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Example 1 Identification of Kochia pollen

[0068] Since the identification and purity of raw materials are extremely important for subsequent allergen diagnosis and therapeutic preparations, in this example, DNA-specific sequence detection and microscopic examination were used to perform double identification and quality control of Kochia 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 of Kochia pollen sample into 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 well, centrifuge at 12000r / min for 5min Take the supernatant into 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] 1. Primer design The primer sequences are as follows: ...

Embodiment 2

[0095] Example 2 Determination of important parameters of Kochia pollen degreasing and leaching key process steps

[0096] 1. Determination of process step parameters for Kochia pollen degreasing

[0097] (1) Degreasing treatment of Kochia pollen (g) and acetone (ml) in a weight-to-volume ratio (w / v) of 1:2. The fat content of different batches (different collection times) of Kochia pollen samples with different degreasing time was tested to determine the optimal degreasing time.

[0098] Haineng SOX500 fat analyzer was used to determine the fat content in Kochia pollen. By Soxhlet extraction and dry weighing, the changes in the weight of pollen before and after extraction were compared to obtain the corresponding fat content. Pollen quality control.

[0099] see the results figure 2 , it can be seen that after the acetone degreasing time is 30 min, the fat content hardly decreases with the extension of the degreasing time, so the degreasing time is determined to be 30 min...

Embodiment 3

[0110] Embodiment 3 Kochia pollen raw material technology

[0111] 1. Pollen collection

[0112] Natural shedding method, collecting Kochia pollen. Dry at room temperature or in a vacuum or fluid bed until the pollen no longer adheres. The dried pollen is passed through a 150-250-mesh sample sieve, preferably a 180-mesh sample sieve in this embodiment.

[0113] 2. Dry

[0114] Spread the pollen in a ventilated and dry place for natural drying, or vacuum drying, or fluidized bed drying for 6-48 hours, until the pollen no longer adheres.

[0115] 3. Degreasing

[0116] The above-obtained pollen (g) and acetone (ml) were subjected to degreasing treatment at a weight-to-volume ratio (w / v) of 1:5 to 1:1. In this embodiment, it is preferably 1:2. After stirring or shaking for 30 minutes, let it stand for stratification, and observe the clarity of the upper layer liquid. Pour off the supernatant, add new acetone, and repeat the degreasing until the supernatant is clear.

[01...

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Abstract

The present invention provides an extract of Kochia pollen allergen, its extract and its preparation method. The extract of Kochia pollen allergen contains the allergenic protein Koc S1', which contains such as SEQ The amino acid sequence shown in ID NO.4. The Kochia pollen allergen extract contains a therapeutically effective dose or a diagnostically effective dose of the Kochia pollen allergen. The infusion solution can also be prepared by collecting, drying, degreasing, extracting, ultrafiltration and concentration, freeze-drying, redissolving and other processes of Kochia pollen. The Kochia pollen allergen extract has the characteristics of high specificity, fully extracted Kochia allergenic protein components, stable total biological potency, long validity period, and good sterility effect; it can be effectively used for skin spots of allergic diseases Prick test diagnosis, in vitro basophil activation test diagnosis and specific immunotherapy can effectively diagnose allergic diseases induced by Kochia pollen and perform specific immunotherapy.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a Kochia pollen allergen extract, an extract thereof and a preparation method thereof. Background technique [0002] People's awareness of allergic diseases began with "hay fever," also known as allergic rhinitis. In 1911, Noon and Freeman first used pollen extract to treat hay fever, and the history of allergy treatment began. At present, allergic diseases are one of the major global health problems. More than 25% of the population in industrialized countries suffers from allergic asthma, allergic rhinoconjunctivitis and allergic dermatitis, of which allergic asthma is the most common. Inhalation of allergenic pollen is the most important factor in causing allergic asthma and other respiratory allergic diseases. VRTA-L.A and other surveys found that nearly 50% of allergic disease patients are allergic to grass pollen. [0003] Ye Xiaoyan's "Analysis of 130 Cases of Chronic Urtic...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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