A kind of ashwagandha pollen allergen extract, its leaching solution and preparation method thereof

An allergen and pollen technology, applied in the biological field, can solve problems such as uncertain components, lower positive rate, and poor stability, and achieve the effects of short cycle, improved stability, and high specificity

Active Publication Date: 2022-08-05
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 reagents are stored for a certain period of time and used for the diagnosis of allergic diseases induced by white wax pollen, the positive rate decreases and the accuracy is low.

Method used

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  • A kind of ashwagandha pollen allergen extract, its leaching solution and preparation method thereof
  • A kind of ashwagandha pollen allergen extract, its leaching solution and preparation method thereof
  • A kind of ashwagandha pollen allergen extract, its leaching solution and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Embodiment 1 Identification of ash 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 dual identification and quality control of the raw materials of the white wax wax.

[0069] 1. Extraction of DNA from P. chinensis pollen

[0070] Tiangen rapid DNA extraction and amplification kit (Tiangen Biochemical KG203) was used.

[0071] Extraction method: Weigh 5 mg of C. chinensis pollen sample into a 1.5 mL centrifuge tube, add 100 μl of Buffer 1, grind the sample thoroughly with a disposable grinding pestle, add 100 μl of Buffer 2, shake and mix, and centrifuge at 12000 r / min for 5 min Then take the supernatant into a new centrifuge tube as a DNA template for future use.

[0072] 2. Primer design and synthesis

[0073] The primer sequences are as follows:

[0074] Fra IT...

Embodiment 2

[0089] Example 2 Determination of the important parameters of the key process steps of degreasing and leaching of the ash pollen

[0090] 1. Determination of the parameters of the degreasing process steps of the ash pollen

[0091](1) Degreasing treatment was carried out with C. chinensis pollen (g) and acetone (ml) at a weight-to-volume ratio (w / v) of 1:2. The fat content of different batches (different collection times) of C. chinensis pollen samples with different defatting times was tested to determine the optimal defatting time.

[0092] Haineng SOX500 fat analyzer was used to determine the fat content in C. chinensis 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.

[0093] 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 degre...

Embodiment 3

[0104] Embodiment 3 Flos ash pollen raw material technology

[0105] 1. Pollen collection

[0106] Natural shedding method to collect ash 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.

[0107] 2. Dry

[0108] 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.

[0109] 3. Degreasing

[0110] 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.

[0111...

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Abstract

The present invention provides the present invention relates to an infusion solution of allergen of C. chinensis pollen, an infusion solution thereof and a preparation method thereof. The allergen extract of C. chinensis pollen contains the amino acid sequence of C. chinensis pollen allergen protein as shown in SEQ ID NO.4. The infusion solution is prepared by the process of collecting, drying, defatting, extracting, ultrafiltration concentration, freeze-drying, redissolving and the like of C. chinensis pollen, and its main components are C. chinensis pollen allergen, glycerin, sodium chloride, phenol, etc. . The allergen infusion solution of C. chinensis pollen has the characteristics of high specificity, sufficient extraction of allergenic protein components of C. chinensis, stable total biological titer, long validity period and good aseptic effect; the original solution can be effectively used for the treatment of allergic diseases. The skin prick test can be used in the diagnosis of in vitro basophil activation test and specific immunotherapy after appropriate dilution.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an infusion solution of allergen of C. chinensis pollen, an infusion solution 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] Fraxinus pennsylvan...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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