CN103613665A - Novel egg yolk antibody and preparation method thereof - Google Patents
Novel egg yolk antibody and preparation method thereof Download PDFInfo
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- CN103613665A CN103613665A CN201310584165.3A CN201310584165A CN103613665A CN 103613665 A CN103613665 A CN 103613665A CN 201310584165 A CN201310584165 A CN 201310584165A CN 103613665 A CN103613665 A CN 103613665A
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- yolk
- yolk antibody
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Abstract
The invention relates to a novel egg yolk antibody and a preparation method thereof, belongs to the field of veterinary biological products, and aims at providing a novel egg yolk antibody capable of effectively preventing and controlling avian influenza, and a refined production technology and use thereof. The egg yolk antibody can be separated from egg white to the maximal extent by adopting an extraction purification method, and is not damaged in purification and concentration processes, and the biological activity of the product is not affected. The method comprises the following steps: immunization of eggs, extraction of the egg yolk antibody, purification of the egg yolk antibody and drying of the egg yolk antibody. The provided egg yolk antibody is mainly applied to detection, prevention and treatment of various avian influenza viruses, and avian influenza immunization failure caused by strain variation.
Description
Technical field
The invention belongs to the moving industry veterinary biologics field of protecting, institute's yolk antibody that provides is mainly for detection of, prevention and treatment multiclass avian influenza, and the bird flu immuning failure causing because of strain variation.
Bird flu is the abbreviation of " avian influenza ", is the bird deadly infectious disease being caused by A type influenza virus, claims again " fowl plague " or " European checken pest ".Clinical main manifestations is acute high lethality, and egg productivity declines, slight respiratory system disease etc.Bird flu is the bird transmissible disease in 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 over more than 100 year, the mankind not yet find effectively prevention and methods for the treatment of.Bird flu or a kind of fowl poultry and people's common sense are sick, and human health is existed to grave danger.
Since in December, 2003, the high pathogenic avian influenza surprise attack Asia being caused by H5N1 avian influenza virus, within the time less than 2 months, 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.Scale that this time bird flu is popular is big, wide, the destructiveness of geographic range in mankind's aviculture history, be all beyond example deeply.Current, bird flu is still active in partial area, the world, continues the gesture of expansion in epidemic situation scope.According to incompletely statistics, from 2003 till now dead the or superseded fowl of ，Jin 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 suffering from the number of bird flu death, has approached 700 people.
Current, develop the medicine of multiple treatment bird flu, and made many feasible prevention and control schemes, and obtained unusual effect.But avian influenza virus, in infectious process, constantly morphs, many effective prevention and control schemes were before lost efficacy.Yolk antibody provided by the present invention is containing Multiple Antibodies, effective to the caused infection of multiple bird flu strain, can reduce the probability that causes prevention and control failure because of strain variation.
Current yolk antibody extracting and purifying method has ethylene glycol extraction method, chloroform extraction method, salting-out process etc., and these method yields are low, foreign protein is many, Product Activity is low.
Summary of the invention
The present invention aims to provide a kind of Novel egg-yolk antibody and refined raw production 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 purifying concentration process can not produce destruction to yolk antibody, does not affect product biological activity.
A kind of Novel egg-yolk antibody and preparation method thereof, its production process is as follows:
1, select high-quality laying hen chicken mattress and purify raising, for producing immune egg, prepare;
2, after laying period starts, to laying hen press certain procedures artificial immunization specific antigen because of or H7, H9 type bird flu attenuated vaccine;
3, collect immune egg and preserve, for extracting yolk antibody, preparing;
4, collect yolk and in the ratio of 1:10, add distilled water and stir 30 minutes, be made into even egg yolk liquid.Regulate egg yolk liquid PH value to 5.5,10 ℃ standing 14 hours, reclaim supernatant liquor;
5, the yolk soil clear liquid containing yolk antibody is carried out to economic benefits and social benefits ultrafiltration and concentration, collecting the concentrated soil of the yolk by recovery of yolk antibody clear liquid filters by the cascade filtration system of 0.45 μ m and 0. 22 μ m, again by filtered liquid with the flow velocity of 4. 5L/min through molecular weight cut-off respectively the economic benefits and social benefits ultrafiltration system of 150KD and 220KD concentrate, solution to be concentrated volume is to reclaim concentrated solution at 1/ 20 o'clock;
6, yolk concentrated solution is carried out to lyophilize, making lyophilized powder filters the yolk soil clear liquid of recovery by the cascade filtration system of 0.45 μ m and 0. 22 μ m, the economic benefits and social benefits ultrafiltration system that again filtered liquid is respectively to 150KD and 220KD with the flow velocity of 4. 5L/min through molecular weight cut-off concentrates, and solution to be concentrated volume is to reclaim concentrated solution at 1/ 20 o'clock;
7, utilize the clinical applicable yolk antibody goods of yolk antibody lyophilized powder preparation to detect anti-body contg in yolk concentrated solution, by 5. 5%, 30% content, add pluronic, Zulkovsky starch respectively, mix rear aseptic subpackaged to sterilizing cillin bottle,-400C lyophilize, makes yolk antibody freeze-dried powder preparation.
Present method is utilized isoelectric point of protein characteristic, by the heavy tactical deployment of troops, yolk antibody is effectively separated with foreign protein, and in 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 producing is high, active strong.
Yolk antibody provided by the present invention is containing Multiple Antibodies, for detection of, prevention and treatment multiclass avian influenza, and made a variation and the bird flu immuning failure that causes by strain.
Embodiment
1: the immunity of laying hen
Select healthy laying hen, carry out head and exempt from H5, H7, H9 type bird flu attenuated vaccine when 170 age in days, vaccination ways is chest muscle multi-point injection.And when exempting from 14 days and 28 days, head carries out respectively reinforced immunological.Within after reinforced immunological 7 days, start to detect yolk antibody level in immune egg, start to collect immune egg after qualified.
2: the extraction of yolk antibody
Use yolk sieve that yolk is separated from immune egg, in the ratio of 1:10, add distilled water and stir 30 minutes, be made into even egg yolk liquid.Regulate egg yolk liquid PH value to 5.0, standing 12 hours of 8 0C, reclaim supernatant liquor.
3: the purifying of yolk antibody
The yolk supernatant liquor of recovery is filtered by the cascade filtration system of 0.45 μ m and 0. 22 μ m, the economic benefits and social benefits ultrafiltration system that again filtered liquid is respectively to 150KD and 220KD with the flow velocity of 4. 5L/min through molecular weight cut-off concentrates, and solution to be concentrated volume is to reclaim concentrated solution at 1/ 20 o'clock.
4: yolk antibody dry
Detect anti-body contg in yolk concentrated solution, add pluronic, Zulkovsky starch respectively by 5.5%, 30% content, mix rear aseptic subpackagedly to sterilizing cillin bottle ,-40 ℃ of lyophilizes, make yolk antibody freeze-dried powder preparation.
Above embodiment is only for the present invention is further illustrated, and scope of the present invention is not subject to the limitation of illustrated embodiment.
Part of test results report: it is 5500 of the sick chickens of mildness bird flu that the trial report of a kind of Novel egg-yolk antibody process for refining and uses thereof is selected clinical diagnosis, 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, ventilates in time, makes a record.Feed, with reference to NRC feeding of broiler standard, is prepared basal diet, and free choice feeding, freely drinks water, unified management.A group is added this yolk antibody freeze-dried powder preparation, in the ratio of 1:100, is watered and drinks; B group, for yellow watt polyoses injection (antiviral I injection liquid), is watered 1kg by every 50ml freely to drink; C group is blank group.
Claims (2)
1. Novel egg-yolk antibody and preparation method thereof, its production process is as follows:
(1), select high-quality laying hen chicken mattress and purify raising, for producing immune egg, prepare;
(2), after laying period starts, to laying hen press certain procedures artificial immunization specific antigen because of or H7, the bird flu of H9 type weaken seedling;
(3), collect immune egg and preserve, for extraction yolk antibody is prepared
(4), collect yolk and in the ratio of 1:10, add distilled water and stir 20 minutes, be made into even egg yolk liquid, regulate egg yolk liquid PH value to 5.5,10 ℃ standing 14 hours, reclaim supernatant liquor;
(5), the yolk soil clear liquid containing yolk antibody is carried out to economic benefits and social benefits ultrafiltration and concentration, collecting the concentrated soil of the yolk by recovery of yolk antibody clear liquid filters by the cascade filtration system of 0.45 μ m and 0. 22 μ m, the economic benefits and social benefits ultrafiltration system that again filtered liquid is respectively to 150KD and 220KD with the flow velocity of 4. 5L/min through molecular weight cut-off concentrates, and solution to be concentrated volume is to reclaim concentrated solution at 1/ 20 o'clock;
(6), yolk concentrated solution is carried out to lyophilize, making lyophilized powder filters the yolk soil clear liquid of recovery by the cascade filtration system of 0.45 μ m and 0. 22 μ m, the economic benefits and social benefits ultrafiltration system that again flow velocity of filtered liquid 4.5L/min is respectively to 150KD and 220KD through molecular weight cut-off concentrates, and solution to be concentrated volume is to reclaim concentrated solution at 1/ 20 o'clock;
(7), utilize the clinical applicable yolk antibody goods of yolk antibody lyophilized powder preparation.
2. according to a kind of Novel egg-yolk antibody described in claim 1, yolk antibody can be made to the clinical applicable formulations such as pulvis, solution, sprays, injection liquid, injection lyophilized powder.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310584165.3A CN103613665A (en) | 2013-11-20 | 2013-11-20 | Novel egg yolk antibody and preparation method thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310584165.3A CN103613665A (en) | 2013-11-20 | 2013-11-20 | Novel egg yolk antibody and preparation method thereof |
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| Publication Number | Publication Date |
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| CN103613665A true CN103613665A (en) | 2014-03-05 |
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| CN201310584165.3A Pending CN103613665A (en) | 2013-11-20 | 2013-11-20 | Novel egg yolk antibody and preparation method thereof |
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| CN (1) | CN103613665A (en) |
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- 2013-11-20 CN CN201310584165.3A patent/CN103613665A/en active Pending
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Application publication date: 20140305 |
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| WD01 | Invention patent application deemed withdrawn after publication |