CN102617732A - Novel egg yolk antibody and preparation method for same - Google Patents
Novel egg yolk antibody and preparation method for same Download PDFInfo
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- CN102617732A CN102617732A CN2012100710105A CN201210071010A CN102617732A CN 102617732 A CN102617732 A CN 102617732A CN 2012100710105 A CN2012100710105 A CN 2012100710105A CN 201210071010 A CN201210071010 A CN 201210071010A CN 102617732 A CN102617732 A CN 102617732A
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- yolk antibody
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Abstract
The invention relates to a novel egg yolk antibody and a preparation method for the same, which belong to the field of biological products for livestock. The invention aims to particularly provide a novel egg yolk antibody for effectively preventing and controlling avian influenza, a refined production process and application. Using an extraction and purification method enables egg yolk antibody to be separated from impurity protein maximally, and a purification and concentration process cannot destroy the egg yolk antibody and affect product biological activity. The method includes the steps of immunization of laying hens, and extraction, purification and drying of the egg yolk antibody. The novel egg yolk antibody is mainly used for detecting, preventing and treating various avian influenza virus infections and avian influenza immunity failure caused by strain variation.
Description
Technical field
The invention belongs to the moving industry veterinary biologics field of protecting, the yolk antibody that provides is mainly used in detection, prevention and treatment multiclass avian influenza, and the bird flu immuning failure that causes because of the strain variation.
Bird flu is the abbreviation of " avian influenza ", is the bird deadly infectious disease that is caused by A type influenza virus, claims again " fowl plague " or " European checken pest ".Clinical acute high lethality, egg productivity decline, the slight respiratory system disease etc. of mainly showing as.Bird flu is the bird transmissible disease in the worldwide, because of the crushing financial loss that aviculture is caused, and is regarded as the No.1 transmissible disease of serious threat poultry farming, and the mankind find effectively to prevent and treat-ment as yet over more than 100 year.Bird flu still is that stud bird poultry and people's common sense are sick, and there is grave danger in human health.
Since in December, 2003; High pathogenic avian influenza by the H5N1 bird flu virus causes is attacked the Asia suddenly; In less than 2 months time, 8 East Asia such as Korea S, Japan, Vietnam, Thailand, Cambodia, Laos, the nations of China and India Nicaea and South Asian nation have been swept across.This time wide, the destructiveness of big, the geographic range of bird flu popular scale all is beyond example on human aviculture history deeply.Current, bird flu is still active in partial area, the world, and the epidemic situation scope is the gesture that continues expansion.According to incompletely statistics, from 2003 till now, only dead the or fowl of eliminating of countries in Asia only sum approximately, 1,500,000,000, the whole world is existing to be surpassed 1000 people and infects bird flu, because of suffering from the dead number of bird flu near 700 people.
Current, develop the medicine of multiple treatment bird flu, and made many feasible prevention and control schemes, and obtained unusual effect.But bird flu virus constantly morphs in infectious process, and many effective prevention and control schemes were before lost efficacy.Yolk antibody provided by the present invention contains multiple antibody, and is effective to the caused infection of multiple bird flu strain, can reduce the probability that causes the prevention and control failure because of the strain variation.
Current yolk antibody extracting and purifying method has ethylene glycol extraction method, chloroform extraction method, sad extraction process, salting-out process etc., and these method yields are low, foreign protein is many, product is active low.
Summary of the invention
The present invention aims to provide a kind of novel yolk antibody and refined raw prodn. art and purposes of effectively prevention and control of fowl influenza.The present invention adopts extracting and purifying method, can to greatest extent yolk antibody be separated from foreign protein, and the purifying concentration process can not produce destruction to yolk antibody, does not influence the product biological activity.
A kind of novel yolk antibody and preparation method thereof, its production process is following:
1, selects high-quality laying hen baby chick and purify raising, prepare for producing immune egg;
2, after laying period begins, to laying hen by certain program artificial immunization specific antigen H5 or H7, H9 type bird flu attenuated vaccine;
3, collect immune egg and preservation, prepare for extracting yolk antibody;
4, collect yolk
Add zero(ppm) water and stirred 30 minutes in the ratio of 1:10, be made into even egg yolk liquid.Regulate egg yolk liquid pH value to 5.0,8 ℃ left standstill 12 hours, reclaimed supernatant;
The yolk supernatant that 5, will contain yolk antibody carries out the economic benefits and social benefits ultrafiltration and concentration, collects yolk antibody and concentrates
With the cascade filtration system filtration of the yolk supernatant that reclaims through 0.45 μ m and 0.22 μ m; The flow velocity of filtered liq with 3.5L/min concentrated through the economic benefits and social benefits ultrafiltration system that molecular weight cut-off is respectively 160KD and 200KD, the solution to be concentrated volume is 1/20 o'clock recovery liquid concentrator again;
6, the yolk liquid concentrator is carried out lyophilize, process lyophilized powder
With the cascade filtration system filtration of the yolk supernatant that reclaims through 0.45 μ m and 0.22 μ m; The flow velocity of filtered liq with 3.5L/min concentrated through the economic benefits and social benefits ultrafiltration system that molecular weight cut-off is respectively 160KD and 200KD, the solution to be concentrated volume is 1/20 o'clock recovery liquid concentrator again;
7, utilize the yolk antibody lyophilized powder to prepare clinical suitable yolk antibody goods
Detect anti-body contg in the yolk liquid concentrator, press 2.5%, 20% content adding pluronic, Zulkovsky starch respectively, mix the aseptic subpackaged sterilization cillin bottle that arrives in back, the yolk antibody freeze-dried powder preparation is processed in-40 ℃ of lyophilizes.
Present method is utilized the isoelectric point of protein characteristic, through settling process yolk antibody is effectively separated with foreign protein, and in the purifying concentration process, does not destroy the space structure of yolk antibody, the biological activity of the yolk antibody that adequately protected.The yolk antibody purity of being produced is high, active strong.
Yolk antibody provided by the present invention contains multiple antibody, and be used for detecting, prevent and treatment multiclass avian influenza, and the bird flu immuning failure that causes by the strain variation.
Embodiment
1: the immunity of laying hen
Select healthy laying hen, when 170 ages in days, carry out head and exempt from H5, H7, H9 type bird flu attenuated vaccine, vaccination ways is the chest muscle multi-point injection.And, head carries out reinforced immunological respectively when exempting from 14 days and 28 days.Begin to detect yolk antibody level in the immune egg behind the reinforced immunological in 7 days, begin to collect immune egg after qualified.
2: the extraction of yolk antibody
Use the yolk sieve that yolk is separated from immune egg, add zero(ppm) water and stirred 30 minutes, be made into even egg yolk liquid in the ratio of 1:10.Regulate egg yolk liquid pH value to 5.0,8 ℃ left standstill 12 hours, reclaimed supernatant.
3: the purifying of yolk antibody
With the cascade filtration system filtration of the yolk supernatant that reclaims through 0.45 μ m and 0.22 μ m; The flow velocity of filtered liq with 3.5L/min concentrated through the economic benefits and social benefits ultrafiltration system that molecular weight cut-off is respectively 160KD and 200KD, the solution to be concentrated volume is 1/20 o'clock recovery liquid concentrator again.
4: the drying of yolk antibody
Detect anti-body contg in the yolk liquid concentrator, press 2.5%, 20% content adding pluronic, Zulkovsky starch respectively, mix the aseptic subpackaged sterilization cillin bottle that arrives in back, the yolk antibody freeze-dried powder preparation is processed in-40 ℃ of lyophilizes.
Above embodiment has been merely the present invention has been further described, and scope of the present invention does not receive the limitation of the embodiment that lifts.
The part of test results report:
The trial report of a kind of novel yolk antibody process for refining and uses thereof
Selecting clinical diagnosis for use is 4200 of the sick chickens of mildness bird flu, and 40 of random chooses are blank the most, and all the other are equally divided into two groups of A, B.Online flat supporting keeps clean, in time ventilates, and carries out record.Feed is prepared basal diet with reference to NRC feeding of broiler standard, and free choice feeding is freely drunk water, unified management.The A group is added this yolk antibody freeze-dried powder preparation, converts water in the ratio of 1:100 and drinks; The B group is converted water 1kg by every 50ml freely to drink for Radix Astragali polysaccharide injection (antiviral I injection liquid); The C group is the blank group.
Table 1 A, B, three groups of treatments of C comparative test result
Can be found out that by table 1 used this kind to contain the sick chicken of the freeze-dried powder preparation treatment of yolk antibody, case fatality rate obviously descended since the 3rd day, food consumption is obviously recovered, four days basic end of medicine for treatment, and effect is superior to control group.Therefore explaining that this kind contains the freeze-dried powder preparation of yolk antibody can enhancing body immunizing power, the treatment bird flu is infected.
Claims (2)
1. novel yolk antibody and preparation method thereof, its production process is following:
(1), select high-quality laying hen baby chick and purify raising, prepare for producing immune egg;
(2), after laying period begins, to laying hen by certain program artificial immunization specific antigen H5 or H7, H9 type bird flu attenuated vaccine;
(3), collect immune egg and preservation, for the extraction yolk antibody is prepared;
(4), collect yolk
Add zero(ppm) water and stirred 30 minutes in the ratio of 1:10, be made into even egg yolk liquid,
Regulate egg yolk liquid pH value to 5.0,8 ℃ left standstill 12 hours, reclaimed supernatant;
(5), the yolk supernatant that will contain yolk antibody carries out the economic benefits and social benefits ultrafiltration and concentration, collect yolk antibody and concentrate
With the cascade filtration system filtration of the yolk supernatant that reclaims through 0.45 μ m and 0.22 μ m; The flow velocity of filtered liq with 3.5L/min concentrated through the economic benefits and social benefits ultrafiltration system that molecular weight cut-off is respectively 160KD and 200KD, the solution to be concentrated volume is 1/20 o'clock recovery liquid concentrator again;
(6), the yolk liquid concentrator is carried out lyophilize, process lyophilized powder
With the cascade filtration system filtration of the yolk supernatant that reclaims through 0.45 μ m and 0.22 μ m; The flow velocity of filtered liq with 3.5L/min concentrated through the economic benefits and social benefits ultrafiltration system that molecular weight cut-off is respectively 160KD and 200KD, the solution to be concentrated volume is 1/20 o'clock recovery liquid concentrator again;
(7), utilize the yolk antibody lyophilized powder to prepare clinical suitable yolk antibody goods.
2. according to the said a kind of novel yolk antibody of claim 1, can yolk antibody be processed clinical suitable formulations such as pulvis, solution, sprays, injection liquid, injection lyophilized powder.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012100710105A CN102617732A (en) | 2012-03-19 | 2012-03-19 | Novel egg yolk antibody and preparation method for same |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012100710105A CN102617732A (en) | 2012-03-19 | 2012-03-19 | Novel egg yolk antibody and preparation method for same |
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| CN102617732A true CN102617732A (en) | 2012-08-01 |
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| CN2012100710105A Pending CN102617732A (en) | 2012-03-19 | 2012-03-19 | Novel egg yolk antibody and preparation method for same |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103694348A (en) * | 2013-12-10 | 2014-04-02 | 刘聚祥 | Preparation and application method for treating swine viral diarrhea biological agent |
| CN103992403A (en) * | 2014-05-23 | 2014-08-20 | 安徽科技学院 | Method for removing lipoprotein and lipid in goose/duck yolk by using freezing method |
| CN106749639A (en) * | 2016-11-23 | 2017-05-31 | 临沂大学 | A kind of new H5 subtype avian influenzas Yolk antibody replaces the method for serum antibody monitoring |
| CN108623679A (en) * | 2018-05-23 | 2018-10-09 | 成都安蒂康生物科技有限公司 | A kind of rapid extracting method of Yolk antibody, the anti influenza product of preparation and its application |
-
2012
- 2012-03-19 CN CN2012100710105A patent/CN102617732A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103694348A (en) * | 2013-12-10 | 2014-04-02 | 刘聚祥 | Preparation and application method for treating swine viral diarrhea biological agent |
| CN103992403A (en) * | 2014-05-23 | 2014-08-20 | 安徽科技学院 | Method for removing lipoprotein and lipid in goose/duck yolk by using freezing method |
| CN106749639A (en) * | 2016-11-23 | 2017-05-31 | 临沂大学 | A kind of new H5 subtype avian influenzas Yolk antibody replaces the method for serum antibody monitoring |
| CN108623679A (en) * | 2018-05-23 | 2018-10-09 | 成都安蒂康生物科技有限公司 | A kind of rapid extracting method of Yolk antibody, the anti influenza product of preparation and its application |
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Application publication date: 20120801 |