Extraction method of dermatophyte cytoplasm protein antigen for preparing egg yolk antibody and egg yolk antibody preparation method

A technology of dermatophyte and protein antigen, which is applied in the preparation of yolk antibody and the extraction of dermatophyte cytoplasmic protein antigen, which can solve the problems of large side effects, slow therapeutic effect, and high recurrence rate

Inactive Publication Date: 2017-01-04
GUANGDONG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present in this new method involves extracting proteins found inside hen's egg shell that can stimulate production of safe Y-like antihelmanum IgG (antithrombopag) by making them more effective at killing yeast fungi called Candida albicans. This results in safer methods for treatments such as prevention or therapy against malaria caused by these organisms.

Problems solved by technology

Technical Problem addressed in this patented technical problem involves finding effective ways to treat molded toenail infestations without causing harmful symptoms such as yeast infections (yeasts) due to their ability to penetrate through layers on the surface layer of the affected parts during growth.

Method used

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  • Extraction method of dermatophyte cytoplasm protein antigen for preparing egg yolk antibody and egg yolk antibody preparation method

Examples

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Embodiment 1

[0028] Example 1: Preparation of dermatophyte cytoplasmic protein antigen

[0029] (1) Cultivate Trichophyton rubrum and Trichophyton mentagrophytes by SDA solid medium, and then inoculate them into liquid medium for mass culture respectively.

[0030] (2) Extraction of cell wall protein: For (1) liquid culture, 10000r / min, 4°C, 10min centrifugation to obtain mycelium, washed repeatedly with PBS, and centrifuged to obtain mycelium; use liquid nitrogen to grind, add PBS to dissolve, add 300mg snail Enzyme breaks the wall; add 100μl DTT, enzymolysis at 37°C for 3h, then 6000r / min, 4°C, 10min to get a precipitate, wash the precipitate repeatedly with pre-cooled PBS (pH=6.4); add cell lysate, blow and mix Afterwards, bathe in ice for 1 hour to completely release the protein in the cytoplasm; centrifuge at 10,000 r / min, 4°C for 20 minutes to obtain the supernatant; dialyze with PBS for three days, and change the dialysate every 12 hours. The clear liquid obtained after dialyzing is...

Embodiment 2

[0031] Example 2: Preparation of egg yolk antibody

[0032] (1) Immunization of laying hens with cytoplasmic protein antigen: select 24 laying hens at the age of 20 weeks; the dermatophyte cytoplasmic protein antigen solution obtained in Example 1 and Freund's complete adjuvant in a volume ratio of 1:1 Mix and emulsify, and inject 0.5ml to each hen by subcutaneous multi-point injection; perform the first booster immunization two weeks after the first immunization, and use an equal volume of Freund's incomplete adjuvant and dermatophyte cytoplasmic protein antigen The solution was mixed and emulsified, and each hen was injected with 0.5ml by subcutaneous multi-point injection, and booster immunization was carried out every 15 days. A total of 4 booster immunizations were performed, and immunized eggs were collected from the first booster immunization, and collected continuously Three-month-old immunized eggs and stored at 4°C;

[0033] (2) Purification of anti-anti-dermatophyt...

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Abstract

The invention belongs to a biological pharmacy technology and discloses an extraction method of a dermatophyte cytoplasm protein antigen for preparing an egg yolk antibody and an egg yolk antibody preparation method. The preparation method specifically comprises the following steps: culturing dermatophyte fungi in an SDA (Sabouraud's dextrose agar) solid culture medium, then inoculating to an SDA liquid culture medium, and performing mass culture; filtering to obtain hyphae, and performing low-temperature grinding with liquid nitrogen; and adding helicase to perform enzymolysis of cell walls, adding the obtained precipitate to a lysis solution, sufficiently reacting, and dialyzing the supernatant with a PBS (phosphate buffer solution) to obtain the dermatophyte cytoplasm protein antigen. Further, a hen is inoculated with the prepared cytoplasm protein used as an antigen to obtain a high-potency antibody, thus providing a new idea for prevention and treatment of dermatophyte diseases.

Description

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Claims

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Application Information

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Owner GUANGDONG UNIV OF TECH
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