CN101717445A - Preparing method of egg yolk antibody - Google Patents
Preparing method of egg yolk antibody Download PDFInfo
- Publication number
- CN101717445A CN101717445A CN200910231294A CN200910231294A CN101717445A CN 101717445 A CN101717445 A CN 101717445A CN 200910231294 A CN200910231294 A CN 200910231294A CN 200910231294 A CN200910231294 A CN 200910231294A CN 101717445 A CN101717445 A CN 101717445A
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- CN
- China
- Prior art keywords
- yolk antibody
- yolk
- antigen
- egg
- mixed solution
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Links
- 210000002969 egg yolk Anatomy 0.000 title claims abstract description 51
- 238000000034 method Methods 0.000 title claims abstract description 11
- 102000002322 Egg Proteins Human genes 0.000 title abstract description 8
- 108010000912 Egg Proteins Proteins 0.000 title abstract description 8
- 235000013345 egg yolk Nutrition 0.000 title abstract 9
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims abstract description 18
- 239000006228 supernatant Substances 0.000 claims abstract description 18
- 239000000427 antigen Substances 0.000 claims abstract description 17
- 102000036639 antigens Human genes 0.000 claims abstract description 17
- 108091007433 antigens Proteins 0.000 claims abstract description 17
- 230000036039 immunity Effects 0.000 claims abstract description 16
- 235000013601 eggs Nutrition 0.000 claims abstract description 14
- 241000287828 Gallus gallus Species 0.000 claims abstract description 10
- 235000011187 glycerol Nutrition 0.000 claims abstract description 9
- 239000002244 precipitate Substances 0.000 claims abstract description 6
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 5
- 238000003756 stirring Methods 0.000 claims abstract description 5
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 claims description 32
- 239000011259 mixed solution Substances 0.000 claims description 20
- 229920003171 Poly (ethylene oxide) Polymers 0.000 claims description 16
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 claims description 16
- -1 polyoxyethylene Polymers 0.000 claims description 16
- 239000002504 physiological saline solution Substances 0.000 claims description 12
- 238000005119 centrifugation Methods 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 6
- 208000010359 Newcastle Disease Diseases 0.000 claims description 5
- 208000015181 infectious disease Diseases 0.000 claims description 5
- 230000002458 infectious effect Effects 0.000 claims description 5
- 239000007924 injection Substances 0.000 claims description 5
- 238000002347 injection Methods 0.000 claims description 5
- 238000001556 precipitation Methods 0.000 claims description 5
- 208000010884 Bursa disease Diseases 0.000 claims description 4
- 241000700605 Viruses Species 0.000 claims description 4
- 239000000839 emulsion Substances 0.000 claims description 4
- 239000008103 glucose Substances 0.000 claims description 4
- 229940031551 inactivated vaccine Drugs 0.000 claims description 4
- 235000013372 meat Nutrition 0.000 claims description 3
- 210000000689 upper leg Anatomy 0.000 claims description 3
- 230000001900 immune effect Effects 0.000 claims description 2
- 239000000047 product Substances 0.000 abstract description 4
- 230000006978 adaptation Effects 0.000 abstract description 2
- 239000004519 grease Substances 0.000 abstract description 2
- 239000007788 liquid Substances 0.000 abstract description 2
- 239000002202 Polyethylene glycol Substances 0.000 abstract 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 abstract 1
- 235000013330 chicken meat Nutrition 0.000 abstract 1
- 239000008121 dextrose Substances 0.000 abstract 1
- 229920001223 polyethylene glycol Polymers 0.000 abstract 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract 1
- 150000002632 lipids Chemical class 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 210000004681 ovum Anatomy 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 238000009461 vacuum packaging Methods 0.000 description 2
- 241000272525 Anas platyrhynchos Species 0.000 description 1
- 208000027312 Bursal disease Diseases 0.000 description 1
- 206010035148 Plague Diseases 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000003452 antibody preparation method Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000005057 refrigeration Methods 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
Landscapes
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The invention discloses a preparing method of egg yolk antibody. The method comprises the following steps of: firstly, collecting eggs laid by layer chickens which are subjected to antigen strengthened immunity; secondly, separating egg yolks from the eggs; thirdly, mixing normal saline containing polyethylene glycol with the egg yolks, uniformly stirring t, and centrifuging to remove precipitate, and taking supernatant; and fourthly, mixing dextrose water containing glycerin with the clear liquid, and centrifuging to remove precipitate, and taking the supernatant which is the egg yolk antibody. By adopting the method, grease in the egg yolks can be thoroughly absorbed, thus greatly reducing the volume of the egg yolk antibody, and improving the purify of the egg yolk antibody. The ratio of purify to mass of the finished product is higher than 95 percent, and the adaptation range of temperature of the prepared egg yolk antibody is wide, and is within 0 to 3 DEG C.
Description
Technical field
The present invention relates to a kind of preparation method of yolk antibody.
Background technology
High-immunity yolk liquid is widely used in treatment of diseases and urgent preventions such as infectious bursal disease, newcastle disease and duck plague, also can extract highly purified diagnostic reagent from yolk.The main component of Ovum Gallus domesticus Flavus is protein and fat, its mass ratio is 1: 2, this shows that the lipid content in the Ovum Gallus domesticus Flavus is sizable, present yolk antibody preparation method is owing to lipid material in the removal yolk is not thorough, lipid material is very big to the influence of yolk antibody, it will cause very thickness of yolk antibody, cause injections difficult when practical application, it is too many to throw the intravital inert matter of fowl into, usually make the swelling of fowl body, stress strengthen, absorption easily forms caseous substance, enlargement or necrosis etc. slowly in the injection site.Have a strong impact on and support the fowl benefit.Make the application of from yolk, extracting antibody be restricted.And general yolk antibody all needs freezing or the refrigeration accumulating, and its suitable temperature is 0-4 ℃.
Summary of the invention
The technical problem to be solved in the present invention is: provide a kind of preparation method of yolk antibody, the yolk antibody purity height that it is made.
In order to solve the problems of the technologies described above, the present invention includes the following step:
(1) collection is through the egg that laying hen produced of antigen reinforced immunological;
(2) isolate yolk in the egg;
(3) mix with yolk with the physiological saline that contains polyoxyethylene glycol and stir, centrifugal removal precipitates, and gets supernatant liquor;
(4) mix centrifugal removal precipitation with supernatant liquor with glycerinated G/W again, get supernatant liquor, this supernatant liquor is the gained yolk antibody.
In order to improve the purity of the yolk antibody of making, the physiological saline that contains polyoxyethylene glycol in the described step (3) be meant the saliferous mass percent be 0.9% physiological saline again with the mixed solution of polyoxyethylene glycol, the polyoxyethylene glycol mass percent is 7% in this mixed solution, and the mass ratio of this mixed solution and yolk is 5: 1.
In order to improve the purity of the yolk antibody of making, the glycerinated G/W of described step (4) be meant contain glucose quality per-cent be 5% G/W again with the mixed solution of glycerine, the mass percent of glycerine is 15% in this mixed solution.
In order to enable to remove fast precipitation, the sedimentary centrifugation rate of centrifugal removal is 4000-8000 rev/min in the described step poly-(3) and (4), and centrifugation time is 3-10 minute.
Has infectious bursa diseases virus antigen in the yolk antibody in order to make, antigen is infectious bursa diseases virus antigen in the described step poly-(1), altogether through twice immunity, and two weeks of interval, the antigen consumption is 1 milliliter of each every chicken, after once two week of immunity the back collect eggs.
Has the newcastle disease oil emulsion inactivated vaccine in the yolk antibody in order to make, antigen is the newcastle disease oil emulsion inactivated vaccine in the described step poly-(1), altogether through three immunity, each immunity 7 days at interval, the antigen consumption is 1 milliliter of each every chicken, adopt the injection of chest thigh meat, egg is collected in last immunity two all backs.
The invention has the beneficial effects as follows: 1, the present invention can thoroughly absorb the grease in the yolk, so just significantly reduced the volume of yolk antibody, improved the purity of yolk antibody, finished product purity mass ratio is up to more than 95%, reduced the consumption of antibody on the one hand, the infringement that when having eliminated injection of antibodies greatly on the other hand the chicken body has been caused has increased the result of use of antibody.
2, the present invention's its thermal adaptation scope of yolk antibody of making is widely 0-30 ℃, has so just guaranteed the activity of antibody greatly, has reduced the freezing link in transportation and the storage process, and is convenient and swift.
3, starting material of the present invention are easily purchased, and are with low cost, simple to operate, not only cost of manufacture low and when using, reduce stress, alleviate side effect, increase the benefit.
Embodiment
Specific embodiment one
This specific embodiment comprises the following steps:
(1) with infectious bursa diseases virus antigen immune laying hen group, altogether through twice immunity, at interval two weeks, the antigen consumption be 1 milliliter of each every chicken, after collection eggs after two weeks of immunity once.
(2) isolate yolk in the egg.
(3) mixes with yolk with the physiological saline that contains polyoxyethylene glycol and stir, centrifugal 10 minutes, 4000 rev/mins of centrifugation rates, removal precipitates, and gets supernatant liquor; The physiological saline that contains polyoxyethylene glycol be meant the saliferous mass percent be 0.9% physiological saline again with the mixed solution of polyoxyethylene glycol, the polyoxyethylene glycol mass percent is 7% in this mixed solution, the mass ratio of this mixed solution and yolk is 5: 1.
(4) mix with supernatant liquor with glycerinated G/W again, centrifugal 5 minutes, 6000 rev/mins of centrifugal speeds, remove precipitation, get supernatant liquor, this supernatant liquor is the gained yolk antibody, glycerinated G/W be meant contain glucose quality per-cent be 5% G/W again with the mixed solution of glycerine, the mass percent of glycerine is 15% in this mixed solution.
(5) finished product is made in vacuum packaging, room temperature preservation.
Specific embodiment two
This specific embodiment comprises the following steps:
(1) with newcastle disease oil emulsion inactivated vaccine immunization laying hen group, altogether through three immunity, immunity is 7 days at interval at every turn, and the antigen consumption is 1 milliliter of each every chicken, adopts chest thigh meat to inject, last two week of immunity back collection egg.
(2) isolate yolk in the egg.
(3) mixes with yolk with the physiological saline that contains polyoxyethylene glycol and stir, centrifugal 3 minutes, 8000 rev/mins of centrifugation rates, removal precipitates, and gets supernatant liquor; The physiological saline that contains polyoxyethylene glycol be meant the saliferous mass percent be 0.9% physiological saline again with the mixed solution of polyoxyethylene glycol, the polyoxyethylene glycol mass percent is 7% in this mixed solution, the mass ratio of this mixed solution and yolk is 5: 1.
(4) mix with supernatant liquor with glycerinated G/W again, centrifugal 8 minutes, 5000 rev/mins of centrifugal speeds, remove precipitation, get supernatant liquor, this supernatant liquor is the gained yolk antibody, glycerinated G/W be meant contain glucose quality per-cent be 5% G/W again with the mixed solution of glycerine, the mass percent of glycerine is 15% in this mixed solution.
(5) finished product is made in vacuum packaging, room temperature preservation.
Claims (6)
1. the making method of a yolk antibody is characterized in that: comprise the following steps:
(1) collection is through the egg that laying hen produced of antigen reinforced immunological;
(2) isolate yolk in the egg;
(3) mix with yolk with the physiological saline that contains polyoxyethylene glycol and stir, centrifugal removal precipitates, and gets supernatant liquor;
(4) mix centrifugal removal precipitation with supernatant liquor with glycerinated G/W again, get supernatant liquor, this supernatant liquor is the gained yolk antibody.
2. the making method of yolk antibody according to claim 1, it is characterized in that: the physiological saline that contains polyoxyethylene glycol in the described step (3) be meant the saliferous mass percent be 0.9% physiological saline again with the mixed solution of polyoxyethylene glycol, the polyoxyethylene glycol mass percent is 7% in this mixed solution, and the mass ratio of this mixed solution and yolk is 5: 1.
3. the making method of yolk antibody according to claim 1 and 2, it is characterized in that: the glycerinated G/W of described step (4) be meant contain glucose quality per-cent be 5% G/W again with the mixed solution of glycerine, the mass percent of glycerine is 15% in this mixed solution.
4. the making method of yolk antibody according to claim 3 is characterized in that: the sedimentary centrifugation rate of centrifugal removal is 4000-8000 rev/min in the described step poly-(3) and (4), and centrifugation time is 3-10 minute.
5. the making method of yolk antibody according to claim 3, it is characterized in that: antigen is infectious bursa diseases virus antigen in the described step poly-(1), altogether through twice immunity, and two weeks of interval, the antigen consumption is 1 milliliter of each every chicken, after once two week of immunity the back collect eggs.
6. the making method of yolk antibody according to claim 3, it is characterized in that: antigen is the newcastle disease oil emulsion inactivated vaccine in the described step poly-(1), altogether through three immunity, each immunity 7 days at interval, the antigen consumption is 1 milliliter of each every chicken, adopt the injection of chest thigh meat, egg is collected in last immunity two all backs.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009102312948A CN101717445B (en) | 2009-12-22 | 2009-12-22 | Preparing method of egg yolk antibody |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009102312948A CN101717445B (en) | 2009-12-22 | 2009-12-22 | Preparing method of egg yolk antibody |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101717445A true CN101717445A (en) | 2010-06-02 |
| CN101717445B CN101717445B (en) | 2012-06-27 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2009102312948A Expired - Fee Related CN101717445B (en) | 2009-12-22 | 2009-12-22 | Preparing method of egg yolk antibody |
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| Country | Link |
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| CN (1) | CN101717445B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102716476A (en) * | 2012-05-31 | 2012-10-10 | 郑州后羿制药有限公司 | Cell inactivated vaccine, egg yolk antibody injection, and preparation method of cell inactivated vaccine |
| CN104208675A (en) * | 2013-09-30 | 2014-12-17 | 郑州后羿制药有限公司 | Mixed freeze-dried powder for preventing chicken viral diseases and preparation method thereof |
| CN112826931A (en) * | 2021-01-06 | 2021-05-25 | 周陶 | Preparation method of anti-HPV and anti-gynecological inflammation composite ovovitellin gel |
-
2009
- 2009-12-22 CN CN2009102312948A patent/CN101717445B/en not_active Expired - Fee Related
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102716476A (en) * | 2012-05-31 | 2012-10-10 | 郑州后羿制药有限公司 | Cell inactivated vaccine, egg yolk antibody injection, and preparation method of cell inactivated vaccine |
| CN102716476B (en) * | 2012-05-31 | 2015-04-22 | 郑州后羿制药有限公司 | Cell inactivated vaccine, egg yolk antibody injection, and preparation method of cell inactivated vaccine |
| CN104208675A (en) * | 2013-09-30 | 2014-12-17 | 郑州后羿制药有限公司 | Mixed freeze-dried powder for preventing chicken viral diseases and preparation method thereof |
| CN104208675B (en) * | 2013-09-30 | 2016-08-17 | 郑州后羿制药有限公司 | A kind of freeze-dried mixed powder of anti-chicken viral diseases and preparation method thereof |
| CN112826931A (en) * | 2021-01-06 | 2021-05-25 | 周陶 | Preparation method of anti-HPV and anti-gynecological inflammation composite ovovitellin gel |
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| Publication number | Publication date |
|---|---|
| CN101717445B (en) | 2012-06-27 |
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| C14 | Grant of patent or utility model | ||
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| C56 | Change in the name or address of the patentee |
Owner name: SHANDONG HUAMU TIANYUAN A + H STOCK CO., LTD. Free format text: FORMER NAME: SHANDONG HUAMUTIANYUAN ANIMAL PRODUCTS CO., LTD. |
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| CP03 | Change of name, title or address |
Address after: 250101, B21, North Building B, environmental protection science park, Lixia District (hi tech Zone), Shandong, Ji'nan Patentee after: SHANDONG HUAMUTIANYUAN A & H STOCK Co.,Ltd. Address before: 250101 B21, North Building B, environmental protection science and technology zone, Licheng hi tech Zone, Licheng District, Shandong, Ji'nan Patentee before: Shandong Huamutianyuan Animal Health Product Co.,Ltd. |
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| EE01 | Entry into force of recordation of patent licensing contract |
Application publication date: 20100602 Assignee: JINAN COMPSUN BIO-TECH Co.,Ltd. Assignor: SHANDONG HUAMUTIANYUAN A & H STOCK Co.,Ltd. Contract record no.: 2013370000007 Denomination of invention: Preparing method of egg yolk antibody Granted publication date: 20120627 License type: Exclusive License Record date: 20130125 |
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| LICC | Enforcement, change and cancellation of record of contracts on the licence for exploitation of a patent or utility model | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
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| CF01 | Termination of patent right due to non-payment of annual fee |