CN104208712A - Chicken infectious bursal disease yolk antibody heatproof freeze-drying protective agent and preparation method thereof - Google Patents
Chicken infectious bursal disease yolk antibody heatproof freeze-drying protective agent and preparation method thereof Download PDFInfo
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- CN104208712A CN104208712A CN201310450261.9A CN201310450261A CN104208712A CN 104208712 A CN104208712 A CN 104208712A CN 201310450261 A CN201310450261 A CN 201310450261A CN 104208712 A CN104208712 A CN 104208712A
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Abstract
The invention discloses a chicken infectious bursal disease yolk antibody heatproof freeze-drying protective agent and a preparation method thereof. The heatproof freeze-drying protective agent is composed of the following components in percentage by weight: 8 to 10% of xylo-oligosaccharide, 2 to 3% of sodium carboxymethylcellulose, 2 to 4% of monosodium glutamate, 3 to 5% of sorbitol, 0.5 to 0.8% of sodium selenite, 2 to 8% of glycerin, and the balance being injection water. The provided protective agent can preserve the chicken infectious bursal disease yolk antibody for 36 months at a temperature of 2 to 8 DEG C, moreover, the shape, activity, and titer of the antibody are not prominently affected during the freeze-drying process and the preservation process, the yolk loss rate is low before and after the freeze-drying, the freeze-drying product has a good anti-aging performance, the speed of the activity degradation of yolk antibody is decelerated, the activity of the pharmaceutical active components can be maintained for a long time, and thus the costs of transportation and preservation are reduced.
Description
Technical field
The invention belongs to veterinary drug technical field, be specifically related to a kind of infectious bursal disease yolk antibody heat-resisting lyophilized protecting agent, also relate to a kind of preparation method of infectious bursal disease yolk antibody heat-resisting lyophilized protecting agent simultaneously.
Background technology
Freeze Drying Technique is biological a kind of important application technology, is usually used in the research and production of biological product, and to keep the biological characteristics of bacterium, seed culture of viruses, stabilate raw material and half-finished biological activity, extend the effect duration of finished product.Freeze drying protectant is the composition giving effective ingredient storage stage protection medicine effect after freezing dry process and lyophilizing, is the key technology in live vaccine or antibody producing, has a great impact for the preservation of vaccine or antibody, transport and use tool.
Veterinary biologics General Requirements cryopreservation, only can about 6 months be preserved under 4 DEG C of conditions, in conjunction with the aquaculture level that China is current, quite a few raiser does not possess the primary condition of cryopreservation biological product, if therefore can not be finished in time at normal temperatures, very easily cause the inefficacy of biological product, bring economic loss to raiser.At present, the freeze drying protectant main component of China's veterinary biologics is skim milk, the sucrose of 5% ~ 10%, the gelatin of 1.5% ~ 3% of 5%, and prescription is simple, and defencive function is poor.On market, the longest protection period of existing freeze drying protectant to yolk antibody is 18 months; have impact on the performance of yolk antibody effect to a certain extent; and the ice crystal that the freeze drying process of infectious bursal disease yolk antibody is formed causes injury in various degree to yolk antibody; directly affect the activity of antibody in dry rear product, and then have influence on the quality of product.
Summary of the invention
The object of this invention is to provide a kind of infectious bursal disease yolk antibody heat-resisting lyophilized protecting agent, extend the pot-life of yolk antibody, effectively keep the curative effect of yolk antibody.
Second object of the present invention is to provide a kind of preparation method of infectious bursal disease yolk antibody heat-resisting lyophilized protecting agent.
In order to realize above object; the technical solution adopted in the present invention is: a kind of infectious bursal disease yolk antibody heat-resisting lyophilized protecting agent; be made up of the component of following percentage by weight: oligomeric xylose 8% ~ 10%, sodium carboxymethyl cellulose 2% ~ 3%, sodium glutamate 2% ~ 4%, sorbitol 3% ~ 5%, sodium selenite 0.5% ~ 0.8%, glycerol 2% ~ 8%, surplus is water for injection.
A preparation method for above-mentioned infectious bursal disease yolk antibody heat-resisting lyophilized protecting agent, comprises the following steps:
1) get 1/3rd of the total consumption of water for injection, add oligomeric xylose, sorbitol, mix homogeneously, obtain mixture A;
2) get 1/3rd of the total consumption of water for injection, add sodium carboxymethyl cellulose, mix homogeneously, obtain mixture B;
3) water for injection under remainder, adds sodium glutamate, sodium selenite, glycerol, mix homogeneously, obtains mixture C;
4) mixture A, mixture B, mixture C mixed and stir, to obtain final product.
Infectious bursal disease yolk antibody heat-resisting lyophilized protecting agent of the present invention, during use, mixes heat-resisting lyophilized protecting agent according to the ratio that volume ratio is 3 ~ 5:1 with infectious bursal disease yolk antibody, after subpackage, carries out lyophilization.
Infectious bursal disease yolk antibody heat-resisting lyophilized protecting agent of the present invention, by oligomeric xylose, sodium carboxymethyl cellulose, sodium glutamate, sorbitol, sodium selenite and glycerol carry out composite, make infectious bursal disease yolk antibody can preserve 36 months under 2 ~ 8 DEG C of conditions, the profile of yolk antibody can be kept in freeze-drying process and preservation process, active, and antibody titer does not have significant change, before and after lyophilizing, yolk antibody loss rate is low, freeze-drying prods has good ageing-resistant performance, slow down the speed of yolk antibody activity decrease, avoid the actual effect of active constituents of medicine in a long time, thus reduce transport, the cost stored.
Detailed description of the invention
Below in conjunction with detailed description of the invention, the present invention is further illustrated.
Embodiment 1
The infectious bursal disease yolk antibody heat-resisting lyophilized protecting agent of the present embodiment; be made up of the component of following percentage by weight: oligomeric xylose 8%, sodium carboxymethyl cellulose 2%, sodium glutamate 2%, sorbitol 3%, sodium selenite 0.5%, glycerol 2%, surplus is water for injection.
A preparation method for above-mentioned infectious bursal disease yolk antibody heat-resisting lyophilized protecting agent, comprises the following steps:
1) get 1/3rd of the total consumption of water for injection, add oligomeric xylose, sorbitol, mix homogeneously, obtain mixture A;
2) get 1/3rd of the total consumption of water for injection, add sodium carboxymethyl cellulose, be heated to 100 DEG C and be incubated 20min, make its mix homogeneously, be cooled to room temperature, obtain mixture B;
3) water for injection under remainder, adds sodium glutamate, sodium selenite, glycerol, mix homogeneously, obtains mixture C;
4) mixture A, mixture B, mixture C mixed and stir, obtaining heat-resisting lyophilized protecting agent.
Mixed according to the ratio that volume ratio is 3:1 with avian infectious bursal disease yolk antibody by the present embodiment gained heat-resisting lyophilized protecting agent, after subpackage, carry out lyophilization, rear sealing, packaging products obtained therefrom can preserve 30 months under 2 ~ 8 DEG C of conditions.
Embodiment 2
The infectious bursal disease yolk antibody heat-resisting lyophilized protecting agent of the present embodiment; be made up of the component of following percentage by weight: oligomeric xylose 9%, sodium carboxymethyl cellulose 2.5%, sodium glutamate 3%, sorbitol 4%, sodium selenite 0.6%, glycerol 8%, surplus is water for injection.
A preparation method for above-mentioned infectious bursal disease yolk antibody heat-resisting lyophilized protecting agent, comprises the following steps:
1) get 1/3rd of the total consumption of water for injection, add oligomeric xylose, sorbitol, mix homogeneously, obtain mixture A;
2) get 1/3rd of the total consumption of water for injection, add sodium carboxymethyl cellulose, be heated to 100 DEG C and be incubated 20min, make its mix homogeneously, be cooled to room temperature, obtain mixture B;
3) water for injection under remainder, adds sodium glutamate, sodium selenite, glycerol, mix homogeneously, obtains mixture C;
4) mixture A, mixture B, mixture C mixed and stir, obtaining heat-resisting lyophilized protecting agent.
Mixed according to the ratio that volume ratio is 4:1 with avian infectious bursal disease yolk antibody by the present embodiment gained heat-resisting lyophilized protecting agent, after subpackage, carry out lyophilization, rear sealing, packaging products obtained therefrom can preserve 36 months under 2 ~ 8 DEG C of conditions.
Embodiment 3
The infectious bursal disease yolk antibody heat-resisting lyophilized protecting agent of the present embodiment; be made up of the component of following percentage by weight: oligomeric xylose 10%, sodium carboxymethyl cellulose 3%, sodium glutamate 4%, sorbitol 5%, sodium selenite 0.8%, glycerol 4%, surplus is water for injection.
A preparation method for above-mentioned infectious bursal disease yolk antibody heat-resisting lyophilized protecting agent, comprises the following steps:
1) get 1/3rd of the total consumption of water for injection, add oligomeric xylose, sorbitol, mix homogeneously, obtain mixture A;
2) get 1/3rd of the total consumption of water for injection, add sodium carboxymethyl cellulose, be heated to 100 DEG C and be incubated 20min, make its mix homogeneously, be cooled to room temperature, obtain mixture B;
3) water for injection under remainder, adds sodium glutamate, sodium selenite, glycerol, mix homogeneously, obtains mixture C;
4) mixture A, mixture B, mixture C mixed and stir, obtaining heat-resisting lyophilized protecting agent.
Mixed according to the ratio that volume ratio is 5:1 with avian infectious bursal disease yolk antibody by the present embodiment gained heat-resisting lyophilized protecting agent, after subpackage, carry out lyophilization, rear sealing, packaging products obtained therefrom can preserve 30 months under 2 ~ 8 DEG C of conditions.
Experimental example
This experimental example adopts the freeze drying protectant preparation infectious bursal disease yolk antibody carry out lyophilization of embodiment 1 ~ 3, preserves under being placed in 2 ~ 8 DEG C, 37 DEG C conditions respectively, detects yolk antibody to tire situation of change in different time.
Experimental technique: regularly taken out by the infectious bursal disease yolk antibody be kept under 2 ~ 8 DEG C, 37 DEG C conditions, indicate plumage part by label, by antibody normal saline dilution to 1 Yu Fen ∕ ml.In 8.0% sodium chloride solution 100ml, add 1g high-quality agar powder, after heating and melting, adding final concentration is that 0.01% thimerosal is anticorrosion, casts gel slab, and press hexagon punching thereon, aperture is 5mm, and pitch-row is 3mm.Test after antibody being made 2 times of serial dilutions, antigen and antibody dosage are with Kong Manwei degree (about 25ul); Then agar plate is put in wet box, hatch 24 ~ 28 hours for 37 DEG C.When positive and negative serum control is set up, the antibody maximum dilution multiple forming white precipitate line with standard antigen is tiring of yolk antibody.
In this experiment, matched group 1 is for using the infectious bursal disease yolk antibody of the common freeze drying protectant preparation of conventional 5% sucrose skimmed milk preparation; Matched group 2 is the infectious bursal disease refine yolk antibody that Shandong company produces; The infectious bursal disease yolk antibody that experimental group 1,2,3 is prepared for utilizing the embodiment of the present application 1,2,3 gained heat-resisting lyophilized protecting agent.
Experimental result is as shown in table 1,2.
Table 1 yolk antibody preserves 6,12,18,24,30,36 months titration results under 2 ~ 8 DEG C of conditions
| Group | Tire in basis | 6 months | 12 months | 18 months | 24 months | 30 months | 36 months |
| Experimental group 1 | 1:128 | 1:128 | 1:128 | 1:128 | 1:128 | 1:128 | 1:64 |
| Experimental group 2 | 1:128 | 1:128 | 1:128 | 1:128 | 1:128 | 1:128 | 1:128 |
| Experimental group 3 | 1:128 | 1:128 | 1:128 | 1:128 | 1:128 | 1:128 | 1:64 |
| Matched group 1 | 1:128 | 1:128 | 1:128 | 1:64 | 1:32 | 1:16 | 1:8 |
| Matched group 2 | 1:128 | 1:128 | 1:128 | 1:128 | 1:64 | 1:32 | 1:16 |
Table 2 yolk antibody preserves 7,14 days titration results under 37 DEG C of conditions
| Group | Tire in basis | 7 days | 14 days |
| Experimental group 1 | 1:128 | 1:128 | 1:64 |
| Experimental group 2 | 1:128 | 1:128 | 1:128 |
| Experimental group 3 | 1:128 | 1:128 | 1:64 |
| Matched group 1 | 1:128 | 1:64 | 1:16 |
| Matched group 2 | 1:128 | 1:32 | 1:8 |
As can be seen from table 1,2; use the infectious bursal disease yolk antibody of the embodiment of the present application 1 ~ 3 gained heat-resisting lyophilized protecting agent preparation; at different temperatures than the good stability that the infectious bursal disease yolk antibody prepared with common freeze drying protectant and commercially available infectious bursal disease refine yolk antibody are tired, the holding time of antibody titer extends.Result of the test illustrates, use the infectious bursal disease yolk antibody of the heat-resisting lyophilized protecting agent of the application preparation to have good thermostability, antibody titer has better stability.
Claims (2)
1. an infectious bursal disease yolk antibody heat-resisting lyophilized protecting agent; it is characterized in that: be made up of the component of following percentage by weight: oligomeric xylose 8% ~ 10%, sodium carboxymethyl cellulose 2% ~ 3%, sodium glutamate 2% ~ 4%, sorbitol 3% ~ 5%, sodium selenite 0.5% ~ 0.8%, glycerol 2% ~ 8%, surplus is water for injection.
2. a preparation method for infectious bursal disease yolk antibody heat-resisting lyophilized protecting agent as claimed in claim 1, is characterized in that: comprise the following steps:
1) get 1/3rd of the total consumption of water for injection, add oligomeric xylose, sorbitol, mix homogeneously, obtain mixture A;
2) get 1/3rd of the total consumption of water for injection, add sodium carboxymethyl cellulose, mix homogeneously, obtain mixture B;
3) water for injection under remainder, adds sodium glutamate, sodium selenite, glycerol, mix homogeneously, obtains mixture C;
4) mixture A, mixture B, mixture C mixed and stir, to obtain final product.
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| CN112852666A (en) * | 2021-01-19 | 2021-05-28 | 新疆河润水业有限责任公司 | Preparation method of microbial agent and microbial fertilizer prepared by adopting microbial agent |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102671207A (en) * | 2012-05-30 | 2012-09-19 | 青岛农业大学 | Newcastle disease and avian infectious bronchitis bigeminal live vaccine heat-resisting freeze-drying protective agent and preparation method |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102671207A (en) * | 2012-05-30 | 2012-09-19 | 青岛农业大学 | Newcastle disease and avian infectious bronchitis bigeminal live vaccine heat-resisting freeze-drying protective agent and preparation method |
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| Title |
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| 杨泽林等: "耐热冻干保护剂及其在兽用冻干活疫苗中的应用", 《四川畜牧兽医》 * |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112852666A (en) * | 2021-01-19 | 2021-05-28 | 新疆河润水业有限责任公司 | Preparation method of microbial agent and microbial fertilizer prepared by adopting microbial agent |
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Address after: 451162 Henan economic comprehensive experimentation area, Zhengzhou port, Xingang Province Patentee after: Henan Hou Yi Industry Group Co., Ltd. Address before: 451162 Xingang, Henan, Zhengzhou, Hong Kong airport on the eastern side of the road on the eastern side of Zhengzhou Patentee before: Zhengzhou Houyi Pharmaceutical Co., Ltd. |
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Granted publication date: 20160817 Termination date: 20200927 |