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Extraction technology of egg yolk immunoglobulins

A technology of egg yolk immunoglobulin and extraction process, applied in immunoglobulin, antibacterial immunoglobulin, immunoglobulin from eggs, etc. The mortality rate and the inability to achieve large-scale production, etc., achieve the effects of easy control of the extraction process, low extraction cost and convenient operation.

Active Publication Date: 2017-02-08
ANHUI ANKE BIOTECHNOLOGY (GRP) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, in this patent, laying hens without normal immunization are used to prepare IgY. We found in early research that in large-scale breeding, laying hens without normal immunization are prone to various poultry diseases, and the mortality rate is extremely high. The mortality rate after four months is as high as 90%, while the mortality rate of immunized healthy laying hens is only 1% after four months
Therefore, the method disclosed in this prior art can only be limited to the laboratory and cannot realize large-scale production

Method used

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  • Extraction technology of egg yolk immunoglobulins

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Acquisition of eggs

[0028] 1) Under aseptic conditions, draw Porphyromonas gingivalis (source to be supplemented) on Columbia blood agar plate by four-section line method, and culture in an anaerobic incubator at 37°C for 4 days until black Colony;

[0029] 2) Inoculate the black colony in 3.7% bovine brain heart infusion liquid medium for anaerobic culture for 3 days, and collect Porphyromonas gingivalis by centrifugation;

[0030] 3) Inactivate Porphyromonas gingivalis with 1% formaldehyde, overnight at 4°C, wash 3 times with sterile PBS buffer, and then wash with a concentration of 1.0×10 11 CFU / ml of inactivated bacteria 1ml and the same amount of Freund's incomplete adjuvant and Freund's complete adjuvant were blown and mixed respectively to form two water-in-oil injections corresponding to the needs.

[0031] 4) Healthy laying hens are immunized 4 times first, and each chicken is injected with 1.0ml each time. The first immunization is an injection ...

Embodiment 2

[0032] Embodiment 2: Extraction of yolk globulin

[0033] 1) Wash and sterilize the eggs, break the shell, remove the egg white and yolk coating, stir the egg yolk evenly, freeze at -20°C for 24 hours, thaw quickly, add 9 times the volume of purified water to dilute and stir for 5 hours, adjust the pH with hydrochloric acid solution to 5.0-5.1, stand overnight at 20-23°C;

[0034]2) Centrifuge the egg yolk dilution at 4°C, take the clear liquid and filter it on a filter column to obtain the filtrate. The filler in the filter column is composed of gauze layers arranged up and down and medical absorbent cotton layers;

[0035] 3) Adopt the ultrafiltration membrane that the molecular weight cut off is 100KD to concentrate the filtrate ultrafiltration 9 times, then use 0.9% sodium chloride solution to replace the ultrafiltration concentrate twice in equal amounts, and replace the gained concentrate according to the ratio of 1ml:0.243g Ammonium sulfate was added to the mixture, fu...

Embodiment 3

[0037] Embodiment 3: Extraction of yolk globulin

[0038] 1) Wash and sterilize the eggs, break the shell, remove the egg white and yolk coating, stir the egg yolk evenly, freeze at -20°C for 48 hours, thaw quickly, add 9 times the volume of purified water to dilute and stir for 5 hours, adjust the pH with hydrochloric acid solution to 5.0-5.1, stand overnight at 20-23°C;

[0039] 2) Centrifuge the egg yolk dilution at 4°C, take the clear liquid and filter it on a filter column to obtain the filtrate. The filler in the filter column is composed of gauze layers arranged up and down and medical absorbent cotton layers;

[0040] 3) Adopt the ultrafiltration membrane that the molecular weight cut off is 100KD to concentrate the filtrate ultrafiltration 9 times, then use 0.9% sodium chloride solution to replace the ultrafiltration concentrate twice in equal amounts, and replace the gained concentrate according to the ratio of 1ml:0.243g Ammonium sulfate was added to the mixture, f...

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Abstract

The invention discloses an extraction technology of egg yolk immunoglobulins, comprising the following steps: a) preparing antigen; b) immunizing laying hen, and collecting eggs; c) uniformly stirring membrane-removed egg yolk, freezing at the temperature of minus 20 DEG C + / - 3 DEG C for at least 24-72 h, rapidly unfreezing, adjusting pH, and standing overnight; d) carrying out centrifugal filtration, carrying out ultrafiltration concentration, carrying out displacement by the use of a sodium chloride solution, carrying out precipitation by the use of ammonium sulfate, and dissolving by the use of sodium chloride; and e) successively carrying out dialysis and centrifugal filtration so as to obtain an egg yolk immunoglobulin liquid. By a series of physical-chemical processes such as centrifugation, filtration, precipitation, dialysis, etc., extraction and purification of egg yolk immunoglobulins are carried out in the invention. In comparison with the prior art, the invention has the following advantages: operation is convenient; extraction cost is low; the extraction technology is easy to control; the extracted egg yolk immunoglobulins can be greatly used for treatment of periodontitis; and the effect is obvious. The extraction technology can be adopted for large-scale industrial production.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to a process for extracting egg yolk immunoglobulin. Background technique [0002] There are mainly three kinds of immunoglobulins in poultry, namely IgG, IgM and IgA. Among them, IgG is the main immunoglobulin of poultry. Because there are many differences between the characteristics of avian IgG and the IgG characteristics of mammals, avian IgG is usually called IgY . When the poultry body is stimulated by a specific antigen, it will produce corresponding specific immune antibodies, and the IgY antibodies produced by the body will be transferred from the blood to the yolk in large quantities, and the antibodies stored in the yolk are called yolk immunoglobulins. Yolk immunoglobulin is a new type of high-efficiency green preparation, which has a good effect on the diagnosis, prevention and treatment of diseases caused by bacteria and viruses. [0003] Poultry eggs (such as chick...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/12C07K16/02C07K1/36C07K1/34C07K1/30
CPCC07K16/02C07K16/1203C07K2317/11
Inventor 宋礼华王荣海李增礼周乐春程婷许培倪晓燕储成风董世建沈毅陈子镭
Owner ANHUI ANKE BIOTECHNOLOGY (GRP) CO LTD
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