CN102813933A - Chicken infectious bursal disease egg yolk antibody cryoprotectant - Google Patents
Chicken infectious bursal disease egg yolk antibody cryoprotectant Download PDFInfo
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- CN102813933A CN102813933A CN2012103170607A CN201210317060A CN102813933A CN 102813933 A CN102813933 A CN 102813933A CN 2012103170607 A CN2012103170607 A CN 2012103170607A CN 201210317060 A CN201210317060 A CN 201210317060A CN 102813933 A CN102813933 A CN 102813933A
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- infectious bursal
- bursal disease
- yolk antibody
- freeze drying
- drying protectant
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Abstract
The invention relates to chicken infectious bursal disease egg yolk antibody cryoprotectant which comprises mannitol, dextran, distilled water solution of glycine and soya peptone. The chicken infectious bursal disease egg yolk antibody cryoprotectant is simple in components, low in cost and easy to prepare. The low-temperature storage difficulty of long-distance transportation and preservation of chicken infectious bursal disease egg yolk antibody is solved. By the cryoprotectant, the chicken infectious bursal disease egg yolk antibody can be preserved for more than 12 months under 2-8 DEG C and be preserved for more than 6 months under 25 DEG C.
Description
Technical field
The invention belongs to birds antibody and preserve technical field, be specifically related to a kind of freeze drying protectant of infectious bursal disease yolk antibody.
Background technology
Infectious bursal disease (Infectious bursal disease, general name IBD) is a kind of acute contagious disease of being encroached on young chicken immune system by infectious bursa of Fabricius virus.Nineteen fifty-seven betides the ridge Boluo of Delaware, USA first, so be called gumboro's disease again.Being dispersed throughout at present the flourishing countries and regions of world's poultry husbandry, also is one of important diseases of China's serious threat poultry husbandry in recent years.This disease mortality rate, mortality on the one hand increases, and causes immunosuppressant on the other hand, and multiple vaccine is descended to the immunne response of chicken, causes immuning failure, causes enormous economic loss.
Yolk antibody is a great discovery of current bioscience research field.Hen with the specific antigen immunity after, can produce corresponding specific antibody, and it is stored in the yolk effectively, the immunoglobulin in the yolk (IgG) is claimed to have another name called Yolk immunoglobulin (IgY) by yolk antibody.Since yolk antibody was found in (1962) such as Williams, IgY drew materials conveniently with it, makes simply, and advantages such as low price are widely used in diagnosis, prevention and the treatment of Animal diseases.Wherein, the infectious bursal disease yolk antibody is exploitation one type of yolk antibody series products early, and in the specific drug use of clinical Chang Zuowei infectious bursal disease, effective percentage is the powerful mean of the clinical infectious bursal disease of control up to more than 90%.
At present, domestic infectious bursal disease yolk antibody preparation all is the liquid dosage form that forms through the dilution packing, and it is big to exist volume, and transportation inconvenience needs cryopreservation and transportation.Be easy to cause the reduction of tiring owing to multigelation and transportation are improper; And lyophilization can make extraction antibody under refrigerated state in advance; Remove moisture and obtain lyophilized formulations without the direct distillation of liquid state, it has avoided the liquid antibody of preserving down to be prone to the shortcoming of inactivation changeableness, makes constant product quality; Volume-diminished is convenient to transportation and long-time the preservation.Freeze drying protectant kind commonly used has polyol, saccharide, protein, polymer, aminoacid, salt and surfactant etc.But in view of livestock and poultry cultivation medication characteristics of development, the freeze drying protectant cost can not be too high, the lyophilizing operation wants simple.Therefore, research and develop a kind of new prescription infectious bursal disease yolk antibody freeze drying protectant simple, with low cost and can prepared in batches and just seem particularly urgent and important.
Summary of the invention
Main purpose of the present invention provides a kind of freeze drying protectant of infectious bursal disease yolk antibody, thereby remedies the deficiency of prior art.
The freeze drying protectant of infectious bursal disease yolk antibody of the present invention is for including the distilled water solution of mannitol, dextran and glycine.
Above-mentioned freeze drying protectant, its each concentration of component mass and size percentage ratio g/ml is: 1-5% mannitol, 1-5% dextran, 0.8-1.2% glycine.
Each concentration of component of above-mentioned freeze drying protectant is preferably: 4% mannitol, 3% dextran, 1% glycine.
Also adding in the above-mentioned freeze drying protectant has soy peptone, and the mass and size percentage ratio g/ml concentration of the soy peptone of adding is 5-10%, is preferably 8%.
With infectious bursal disease yolk antibody and above-mentioned freeze drying protectant with volume ratio 7:1 mixed, as infectious bursal disease yolk antibody lyophilizing liquid; Lyophilizing liquid was cooled to-40 ℃ and kept 4 hours in 1 hour; In 1 hour, goods are risen to-8 ℃ and kept 14 hours from-40 ℃ then; 0.5 hour goods are risen to 28 ℃ and keep the preparation of accomplishing infectious bursal disease yolk antibody lyophilized formulations in 10 hours from-8 ℃.
Freeze drying protectant component of the present invention is simple, with low cost, be easy to preparation, solved the long-distance transport of infectious bursal disease yolk antibody, the low temperature storage difficult problem of preservation.Result of use shows that the frozen-dried protective agent prescription can make the infectious bursal disease yolk antibody preserve down more than 12 months at 2-8 ℃, preserves more than 6 months down, has good effect for 25 ℃.
The specific embodiment
Further describe the present invention below in conjunction with specific embodiment, but these instances only are exemplary, conventional method of the present invention are not constituted any restriction.It will be understood by those skilled in the art that and down can make amendment with form or replace without departing from the spirit and scope of the present invention, but these modifications and replacing all fall in the protection domain of the present invention the details of technical scheme of the present invention.
Embodiment 1: the freeze drying protectant preparation of infectious bursal disease yolk antibody
In the distilled water solution of 100ml, add the mannitol of 4g, the dextran of 3g, the glycosides propylhomoserin of 1g, behind the mix homogeneously, transfer pH value to 7.2～7.4, after sterilization treatment, 2～8 ℃ of preservations are subsequent use.
Embodiment 2: added soy peptone
In the distilled water solution of 100ml, add the mannitol of 4g, the dextran of 3g, the glycosides propylhomoserin of 1g, add the soy peptone of 8g again, transfer pH value to 7.2～7.4,2～8 ℃ of preservations are subsequent use after sterilization treatment.
Embodiment 3: the lyophilizing of infectious bursal disease yolk antibody
With the freeze drying protectant of infectious bursal disease yolk antibody and the foregoing description 1 or embodiment 2 with the 7:1 mixed, as infectious bursal disease yolk antibody lyophilizing liquid.The pre-freeze stage: lyophilizing liquid room temperature inlet, goods were reduced in 1 hour-40 ℃ and kept 4 hours.Sublimation stage: goods were risen to-8 ℃ and kept 14 hours from-40 ℃ in 1 hour.The adsorption stripping and dry stage: goods were risen to 28 ℃ and kept 10 hours, omnidistance 30.5 hours of lyophilizing from-8 ℃ in 0.5 hour.
The lyophilized formulations of infectious bursal disease yolk antibody after the lyophilizing is placed 2-8 ℃ and 25 ℃ of environment preservations down, the character of routine observation antibody, residual moisture and effectiveness respectively.Character, residual moisture detection method are referring to existing " veterinary biologics inspection procedure ", and antibody titer adopts two-way agar gel diffusion test (AGP):
Punching: the agar plate for preparing is punched with card punch by template, and choose the agar in the hole.1 hole, center, 6 holes on every side, aperture 3 mm, pitch-row 4 mm are at the bottom of fusing agar perforations adding.
Application of sample: medium pore adds the standard agp antigen, and the hole adds the infectious bursal disease yolk antibody to be checked of 2 times of serial dilutions respectively on every side, each hole application of sample 25 μ l.Agar plate behind the application of sample is put into the box that is filled with wet gauze, put 20-25 ℃ of room temperature or 37 ℃ of incubators, 24 h just declare, and 72 h declare eventually.
The result judges: precipitation line also occurs between infectious bursal disease yolk antibody to be checked and the antigen hole, and match with standard positive serum precipitation line end, promptly be judged to the positive.When no precipitation line occurs between infectious bursal disease yolk antibody to be checked and the antigen hole, promptly be judged to feminine gender.When forming clear precipitation line between high dilution hole of infectious bursal disease yolk antibody to be checked and the antigen hole, promptly be judged to fine jade and expand and tire.
The result: preserve 12 months, 25 ℃ preservations 6 months with 2-8 ℃ after the lyophilizing of infectious bursal disease yolk antibody, it is stable that the antibody physical behavior keeps, and is the loose agglomerate of little yellow, is prone to separate with the bottle wall; Residual moisture is about 2.32%; Antibody titer is more than the 1:128, does not fall as follows.
Table 1 infectious bursal disease yolk antibody lyophilized formulations assay
Annotate: lyophilizing 1: freeze drying protectant is the soybean protein-containing peptone not; Lyophilizing 2: freeze drying protectant adds 8% soy peptone
Added soy peptone in the protective agent of the present invention, can more effectively prevent during preservation that antibody titer from reducing, and makes the renewal of preservation.Vegetable endogenous binding protein matter is more safer than animal-based protein matter simultaneously, has avoided potential animal derived pathogen contamination antibody.
Claims (7)
1. an infectious bursal disease yolk antibody freeze drying protectant is characterized in that, described freeze drying protectant is the distilled water solution that includes mannitol, dextran and glycine.
2. infectious bursal disease yolk antibody freeze drying protectant as claimed in claim 1 is characterized in that the concentration of described mannitol, dextran and glycine is respectively 1-5%, 1-5% and 0.8-1.2%.
3. infectious bursal disease yolk antibody freeze drying protectant as claimed in claim 2 is characterized in that the concentration of described mannitol, dextran and glycine is respectively 4%, 3% and 1%.
4. the described freeze drying protectant of claim 1 is characterized in that, also adding has soy peptone.
5. freeze drying protectant as claimed in claim 4, the concentration that it is characterized in that described soy peptone is 5-10%.
6. freeze drying protectant as claimed in claim 5, the concentration that it is characterized in that described soy peptone is 8%.
7. infectious bursal disease yolk antibody lyophilized formulations; Be with volume ratio 7:1 mixed, as infectious bursal disease yolk antibody lyophilizing liquid with infectious bursal disease yolk antibody and claim 1 or the described freeze drying protectant of claim 4; Lyophilizing liquid was cooled to-40 ℃ and kept 4 hours in 1 hour; In 1 hour, goods are risen to-8 ℃ and kept 14 hours from-40 ℃ then; The adsorption stripping and dry stage: goods were risen to 28 ℃ and keep the preparation of accomplishing infectious bursal disease yolk antibody lyophilized formulations in 10 hours from-8 ℃ in 0.5 hour.
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| Application Number | Priority Date | Filing Date | Title |
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| CN2012103170607A CN102813933A (en) | 2012-08-30 | 2012-08-30 | Chicken infectious bursal disease egg yolk antibody cryoprotectant |
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| CN2012103170607A CN102813933A (en) | 2012-08-30 | 2012-08-30 | Chicken infectious bursal disease egg yolk antibody cryoprotectant |
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| CN1846785A (en) * | 2005-04-05 | 2006-10-18 | 北京诺思兰德生物技术有限责任公司 | Freeze dried composition containing genetic engineering fusion protein |
| CN1893973A (en) * | 2003-12-17 | 2007-01-10 | 惠氏公司 | Methods for porducing storage stable viruses and immunogenic compositions thereof |
| CN101113176A (en) * | 2006-07-28 | 2008-01-30 | 洛阳普莱柯生物工程有限公司 | Method for preparing infectious chicken Fabricius bursa refined yolk cryodesiccation antibody |
| CN101361717A (en) * | 2007-08-08 | 2009-02-11 | 北京赛生药业有限公司 | Batroxobin freeze-dry powder injection and preparation method thereof |
| CN101940586A (en) * | 2010-08-27 | 2011-01-12 | 北京中海康医药科技发展有限公司 | High safety polyene phosphatidyl choline for injection and preparation method thereof |
| CN102228687A (en) * | 2011-06-24 | 2011-11-02 | 浙江普康生物技术股份有限公司 | Freeze-dried live attenuated hepatitis A vaccine not containing gelatin or human albumin protective agent and preparation method for freeze-dried live attenuated hepatitis A vaccine |
| CN102657870A (en) * | 2012-06-04 | 2012-09-12 | 长春祈健生物制品有限公司 | Vaccine cryoprotectant without composition of gelatin and human albumin |
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2012
- 2012-08-30 CN CN2012103170607A patent/CN102813933A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1260171A (en) * | 2000-01-25 | 2000-07-19 | 长春长生基因药业股份有限公司 | Recombined alpha-type interferon freeze-drying protective agent |
| CN1893973A (en) * | 2003-12-17 | 2007-01-10 | 惠氏公司 | Methods for porducing storage stable viruses and immunogenic compositions thereof |
| CN1846785A (en) * | 2005-04-05 | 2006-10-18 | 北京诺思兰德生物技术有限责任公司 | Freeze dried composition containing genetic engineering fusion protein |
| CN101113176A (en) * | 2006-07-28 | 2008-01-30 | 洛阳普莱柯生物工程有限公司 | Method for preparing infectious chicken Fabricius bursa refined yolk cryodesiccation antibody |
| CN101361717A (en) * | 2007-08-08 | 2009-02-11 | 北京赛生药业有限公司 | Batroxobin freeze-dry powder injection and preparation method thereof |
| CN101940586A (en) * | 2010-08-27 | 2011-01-12 | 北京中海康医药科技发展有限公司 | High safety polyene phosphatidyl choline for injection and preparation method thereof |
| CN102228687A (en) * | 2011-06-24 | 2011-11-02 | 浙江普康生物技术股份有限公司 | Freeze-dried live attenuated hepatitis A vaccine not containing gelatin or human albumin protective agent and preparation method for freeze-dried live attenuated hepatitis A vaccine |
| CN102657870A (en) * | 2012-06-04 | 2012-09-12 | 长春祈健生物制品有限公司 | Vaccine cryoprotectant without composition of gelatin and human albumin |
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Address after: 266111 Shandong city of Qingdao province Chengyang Qingda Industrial Park, the first northbound dual Applicant after: QINGDAO VLAND BIOLOGICAL CO., LTD. Applicant after: Qingdao Boite Biopharmaceutical Co., Ltd. Address before: 266061 Shandong city of Qingdao province high tech park of Laoshan district by the No. 29 Shandong Road, building 6F high speed Applicant before: Qingdao KDN Pharmaceutical Co., Ltd. Applicant before: Qingdao Boite Biopharmaceutical Co., Ltd. |
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Application publication date: 20121212 |