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A kind of preparation method of Periplaneta americana allergen preparation

A cockroach and allergen technology, applied in allergic diseases, allergen antigen components, respiratory diseases, etc., to achieve the effect of small error and good repeatability

Active Publication Date: 2016-08-17
何韶衡
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, so far, there is no published preparation process of Periplaneta americana, and the allergen preparation of Periplaneta americana has a large clinical demand, so it is necessary to develop a new generation of standardized Periplaneta americana allergen preparation

Method used

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  • A kind of preparation method of Periplaneta americana allergen preparation

Examples

Experimental program
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Effect test

Embodiment 1

[0021] A kind of preparation method of Periplaneta americana allergen preparation, comprises the steps:

[0022] (1) Periplaneta americana was inactivated in a -80°C refrigerator; the intestinal tract was removed under an operating microscope; cleaned with 75% ethanol, soaked for 15 minutes, air-dried and placed in a -80°C refrigerator for future use;

[0023] (2) Get an appropriate amount of freezing Periplaneta americana and grind it into powder in liquid nitrogen;

[0024] (3) After degreasing with acetone for 2 times, place it in a -80°C refrigerator to freeze, and then lyophilize it into powder in a vacuum freeze dryer;

[0025] (4) Weigh 1 g of Periplaneta americana powder, add 20 L of extract, mix well on a vortex shaker, place on a shaker at 4°C for extraction for 0.5 hour, then centrifuge at 12,000 g at a speed of 4°C for 30 min, Aspirate the supernatant, which is the Periplaneta americana allergen extract.

[0026] (5) Use specific Per a1, Per a2, Per a3, Per a4, P...

Embodiment 2

[0031] A kind of preparation method of Periplaneta americana allergen preparation, comprises the steps:

[0032] (1) Periplaneta americana was inactivated in a -80°C refrigerator; the intestinal tract was removed under an operating microscope; cleaned with 75% ethanol, soaked for 15 minutes, air-dried and stored in a -80°C refrigerator for later use;

[0033] (2) Get an appropriate amount of freezing Periplaneta americana and grind it into powder in liquid nitrogen;

[0034] (3) After degreasing with acetone for 2 times, place it in a -80°C refrigerator to freeze, and then lyophilize it into powder in a vacuum freeze dryer;

[0035] (4) Weigh 1 g of Periplaneta americana powder, add 20 L of extract, mix well on a vortex shaker, place on a shaker at 4°C for extraction for 2 hours, then centrifuge at 12,000 g at a speed of 4°C for 30 min, Aspirate the supernatant, which is the Periplaneta americana allergen extract.

[0036] (5) identify the main allergens in the crude extract...

Embodiment 3

[0041] A kind of preparation method of Periplaneta americana allergen preparation, comprises the steps:

[0042] (1) Periplaneta americana was inactivated in a -80°C refrigerator; the intestinal tract was removed under an operating microscope; cleaned with 75% ethanol, soaked for 15 minutes, air-dried and placed in a -80°C refrigerator for future use;

[0043] (2) Get an appropriate amount of freezing Periplaneta americana and grind it into powder in liquid nitrogen;

[0044] (3) After degreasing with acetone for 2 times, place it in a -80°C refrigerator to freeze, and then lyophilize it into powder in a vacuum freeze dryer;

[0045] (4) Weigh 1 g of Periplaneta americana powder, add 20 L of extract, mix well on a vortex shaker, place on a shaker at 4°C for extraction for 4 hours, then centrifuge at 12,000 g at a speed of 4°C for 30 min, Aspirate the supernatant, which is the Periplaneta americana allergen extract.

[0046] (5) Use specific Per a1, Per a2, Per a3, Per a4, Pe...

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Abstract

The present invention provides a periplaneta americana allergen preparation production method, which comprises: (1) inactivating periplaneta americana in a refrigerator, removing intestinal tracts, washing with 75% ethanol, carrying out air-drying, and placing in the refrigerator so as to be spare; (2) taking an appropriate amount of the frozen periplaneta americana, and grinding into powder in liquid nitrogen; (3) degreasing with acetone, placing into the refrigerator to freeze, and carrying out freeze-drying in a vacuum freeze dryer to obtain powder; (4) weighing the periplaneta americana powder, adding an extraction solution, carrying out centrifugation, and sucking the supernatant, wherein the supernatant is a periplaneta americana allergen extract; (5) adopting specific antibodies to identify the content and the proportion of the main allergen in the periplaneta americana allergen crude extract; and (6) according to the identification result in the step (5), adding recombinant protein expressed by yeast or insects. With the periplaneta americana allergen crude extract extracted by using the method, the main allergen protein of the periplaneta americana can be obtained to the maximal degree, the extract has good repeatability, and the error is less than 30%.

Description

technical field [0001] The invention relates to the technical field of preparation of allergen preparations, in particular to a preparation method of Periplaneta americana allergen preparations. Background technique [0002] Allergic diseases are one of the major human diseases. Its incidence rate is currently estimated to account for about 22% of the world's population, and is increasing at a rate of more than 1% per year, and the incidence rate of children is the most obvious increase. Its harmfulness cannot be simply measured by its mortality rate, but the economic burden caused by the high attack frequency of the disease should be considered. Because allergic diseases mostly occur in young people, their impact on human society and economy is even more serious. In the process of industrialization transformation in my country, its incidence rate has increased significantly, seriously affecting the quality of life and work efficiency of the affected population in my count...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/35A61P37/08A61P11/06
Inventor 何韶衡
Owner 何韶衡
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