CN103102410B - Method for separating anti-rotavirus immunoglobulin from chicken egg - Google Patents
Method for separating anti-rotavirus immunoglobulin from chicken egg Download PDFInfo
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- CN103102410B CN103102410B CN201310023842.4A CN201310023842A CN103102410B CN 103102410 B CN103102410 B CN 103102410B CN 201310023842 A CN201310023842 A CN 201310023842A CN 103102410 B CN103102410 B CN 103102410B
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- egg white
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- ovum gallinaceum
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Abstract
The invention provides a method for separating anti-rotavirus immunoglobulin from a chicken egg. The method comprises the process steps of: transforming the chicken egg at the constant temperature, breaking the egg, adding water for dilution, performing first sedimentation and performing second sedimentation. The adoption of the method disclosed by the invention for separating the anti-rotavirus immunoglobulin from the chicken egg inoculated with rotavirus has the characteristics of simple process, few equipment, low construction cost, capability of ensuring quality and high separation efficiency. The method is suitable for separating the anti-rotavirus immunoglobulin from the chicken egg after the chicken egg is inoculated with the rotavirus.
Description
Technical field
What the present invention proposed is the extracting method of biochemical field material, specifically rotavirus resisting immune globulin separation method in ovum gallinaceum.
Background technology
After rotavirus infection human body, can damage children's digestive system, occur seasonal diarrhoea, even unhealthful level or cause dehydration dead.In order to prepare the antibody of anti-rotavirus, conventionally the method adopting is: rotavirus is injected in the body of chicken, through after a period of time, chicken produces antibody to rotavirus, antibody accumulates in interior each organ of body of chicken, and also accumulate in ovum gallinaceum, want to obtain rotavirus resisting immune globulin, just need to from ovum gallinaceum, extract.Before the present invention proposes, rotavirus resisting immune globulin separates or separates by whizzer through gel chromatographic columns, wherein, gel chromatography column separation process complexity, easily pollute, and extraction efficiency is low, and although whizzer separation separation efficiency is high, but DNA purity is low, loss is serious, and whizzer is not easy to clean.Therefore improving rotavirus resisting immune globulin quality and efficiency is the key issue that needs solution.
Summary of the invention
In order to improve quality and the efficiency of anti-rotavirus, the present invention proposes rotavirus resisting immune globulin separation method in a kind of ovum gallinaceum.The method is carried out constant temperature conversion by the ovum gallinaceum that inoculation rotavirus is produced, and then carries out gradation sedimentation, separates the immunoglobulin (Ig) of anti-rotavirus from egg white liquid.Solve the technical problem of rotavirus resisting immune globulin separation and Extraction in ovum gallinaceum.
The scheme that technical solution problem of the present invention adopts is:
1, ovum gallinaceum constant temperature transforms: ovum gallinaceum departs from after parent, concentrates in container, under 18～25 DEG C of temperature condition, transforms 120 hours.Make the antibody immune globulin molecule propagation in ovum gallinaceum.
2, broken ovum thin up:
The ovum gallinaceum transforming through constant temperature is broken, taken out yolk and egg white, the yolk of one times of quality and egg white are joined in the deionized water of five times of quality, become ovum gallinaceum liquid.
2, sedimentation for the first time: ovum gallinaceum liquid is injected in container, sedimentation at normal temperatures 24 hours, initial gross separation goes out yolk throw out and egg white liquid, and egg white liquid is toppled over to separation, collects egg white liquid, removes yolk throw out.
3, sedimentation for the second time: the egg white liquid through sedimentation is for the first time continued to sedimentation 48 hours at normal temperatures, egg white liquid is divided into three layers, the first layer is water, topple over and separate, the second layer is the egg white liquid that is enriched with rotavirus resisting immune globulin, pour out as rotavirus resisting immune globulin liquid, and bottom is Residual yolk and heavy impurity, discards.
Positively effect, adopts method of the present invention to separate rotavirus resisting immune globulin from the ovum gallinaceum of inoculation rotavirus, has process simple, and equipment used is few, and structure expense is low, and quality can ensure, the feature that separation efficiency is high.The suitable method application that separates rotavirus resisting immune globulin from the ovum gallinaceum inoculation rotavirus.
Embodiment
1, ovum gallinaceum constant temperature transforms: ovum gallinaceum departs from after parent, concentrates in container, under 18～25 DEG C of temperature condition, transforms 120 hours.Make the antibody immune globulin molecule propagation in ovum gallinaceum.
2, broken ovum thin up:
The ovum gallinaceum transforming through constant temperature is broken, taken out yolk and egg white, the yolk of one times of quality and egg white are joined in the deionized water of five times of quality, become ovum gallinaceum liquid.
2, sedimentation for the first time: ovum gallinaceum liquid is injected in container, sedimentation at normal temperatures 24 hours, initial gross separation goes out yolk throw out and egg white liquid, and egg white liquid is toppled over to separation, collects egg white liquid, removes yolk throw out.
3, sedimentation for the second time: the egg white liquid through sedimentation is for the first time continued to sedimentation 48 hours at normal temperatures, egg white liquid is divided into three layers, the first layer is water, topple over and separate, the second layer is the egg white liquid that is enriched with rotavirus resisting immune globulin, pour out as rotavirus resisting immune globulin liquid, and bottom is Residual yolk and heavy impurity, discards.
Can collect more than 95% rotavirus resisting immune globulin raw material in ovum gallinaceum by twice precipitation, compare with whizzer separation method with traditional gel chromatographic columns, can save facility investment more than 80%, and, settlement separate thorough, constant product quality, compounding technology requirement, working method is simple, and production efficiency is high.
Separation method contrast table
| Separation method: | Yield | Elapsed time | Product content |
| Gel chromatographic columns partition method | 80% | 5 days | 90% |
| Whizzer partition method | 75% | 2 days | 85% |
| Settling process | 93% | 3 days | 98% |
Claims (1)
1. rotavirus resisting immune globulin separation method in ovum gallinaceum, 1) conversion of ovum gallinaceum constant temperature: ovum gallinaceum departs from after parent, concentrates in container, under 18～25 DEG C of temperature condition, transforms 120 hours, makes the antibody immune globulin molecule propagation in ovum gallinaceum;
2), broken ovum thin up: the ovum gallinaceum transforming through constant temperature is broken, taken out yolk and egg white, the yolk of one times of quality and egg white are joined in the deionized water of five times of quality, become ovum gallinaceum liquid;
It is characterized in that:
Ovum gallinaceum, under 18～25 DEG C of temperature condition, transforms 120 hours;
The yolk of one times of quality and egg white are joined in the deionized water of five times of quality, become ovum gallinaceum liquid;
Sedimentation for the first time: ovum gallinaceum liquid is injected in container, sedimentation at normal temperatures 24 hours, initial gross separation goes out yolk throw out and egg white liquid, and egg white liquid is toppled over to separation, collects egg white liquid, removes yolk throw out;
Sedimentation for the second time: the egg white liquid through sedimentation is for the first time continued to sedimentation 48 hours at normal temperatures, egg white liquid is divided into three layers, the first layer is water, topple over and separate, the second layer is the egg white liquid that is enriched with rotavirus resisting immune globulin, pour out as rotavirus resisting immune globulin liquid, and bottom is Residual yolk and heavy impurity, discards.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310023842.4A CN103102410B (en) | 2013-01-23 | 2013-01-23 | Method for separating anti-rotavirus immunoglobulin from chicken egg |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310023842.4A CN103102410B (en) | 2013-01-23 | 2013-01-23 | Method for separating anti-rotavirus immunoglobulin from chicken egg |
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| CN103102410A CN103102410A (en) | 2013-05-15 |
| CN103102410B true CN103102410B (en) | 2014-11-05 |
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| CN201310023842.4A Active CN103102410B (en) | 2013-01-23 | 2013-01-23 | Method for separating anti-rotavirus immunoglobulin from chicken egg |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1417232A (en) * | 2002-12-06 | 2003-05-14 | 孙亮 | Production process of extracting antibody immune globulin from hen's egg |
| CN1935841A (en) * | 2006-09-04 | 2007-03-28 | 天津大学 | Anti hepatitis B virus chicken yolk immune globulin IgY and its separating method |
| CN101955532A (en) * | 2010-10-21 | 2011-01-26 | 郑州后羿制药有限公司 | Method for extracting chicken egg yolk antibodies against infectious bursal disease |
| WO2011107883A1 (en) * | 2010-03-04 | 2011-09-09 | Novartis Ag | Avian group d rotavirus |
| CN102276718A (en) * | 2011-07-26 | 2011-12-14 | 广州格雷特生物科技有限公司 | Process for extracting yolk immunoglobulin by water-caprylic acid two-step method |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101948538A (en) * | 2010-10-19 | 2011-01-19 | 郑州后羿制药有限公司 | Method for refining chicken egg yolk antibodies against Escherichia coli |
| CN101948537B (en) * | 2010-10-21 | 2012-12-19 | 郑州后羿制药有限公司 | Method for extracting chicken yolk antibody against infectious bursal disease |
| CN102526727B (en) * | 2012-01-04 | 2014-05-07 | 河北科星药业有限公司 | Anti-bird-flue egg yolk antibody injection and application thereof |
| CN102702347A (en) * | 2012-05-17 | 2012-10-03 | 天津宝迪农业科技股份有限公司 | Method for extracting immunoglobulin G from swine blood |
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2013
- 2013-01-23 CN CN201310023842.4A patent/CN103102410B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1417232A (en) * | 2002-12-06 | 2003-05-14 | 孙亮 | Production process of extracting antibody immune globulin from hen's egg |
| CN1935841A (en) * | 2006-09-04 | 2007-03-28 | 天津大学 | Anti hepatitis B virus chicken yolk immune globulin IgY and its separating method |
| WO2011107883A1 (en) * | 2010-03-04 | 2011-09-09 | Novartis Ag | Avian group d rotavirus |
| CN101955532A (en) * | 2010-10-21 | 2011-01-26 | 郑州后羿制药有限公司 | Method for extracting chicken egg yolk antibodies against infectious bursal disease |
| CN102276718A (en) * | 2011-07-26 | 2011-12-14 | 广州格雷特生物科技有限公司 | Process for extracting yolk immunoglobulin by water-caprylic acid two-step method |
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| CN103102410A (en) | 2013-05-15 |
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