CN104208712B - A kind of infectious bursal disease yolk antibody heat-resisting lyophilized protecting agent and preparation method thereof - Google Patents
A kind of infectious bursal disease yolk antibody heat-resisting lyophilized protecting agent and preparation method thereof Download PDFInfo
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- CN104208712B CN104208712B CN201310450261.9A CN201310450261A CN104208712B CN 104208712 B CN104208712 B CN 104208712B CN 201310450261 A CN201310450261 A CN 201310450261A CN 104208712 B CN104208712 B CN 104208712B
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- yolk antibody
- bursal disease
- infectious bursal
- protecting agent
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Abstract
The invention discloses a kind of infectious bursal disease yolk antibody heat-resisting lyophilized protecting agent and preparation method thereof; this heat-resisting lyophilized protecting agent consists of the following components in 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%, and surplus is water for injection.The infectious bursal disease yolk antibody heat-resisting lyophilized protecting agent of the present invention; make infectious bursal disease yolk antibody can preserve 36 months under the conditions of 2～8 DEG C; the profile of yolk antibody, activity can be kept during freeze-drying process and preservation; and antibody titer does not has 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.
Description
Technical field
The invention belongs to veterinary drug technical field, be specifically related to a kind of infectious bursal disease yolk antibody heat resistant freeze drying protection
Agent, also relates to the preparation method of a kind of infectious bursal disease yolk antibody heat-resisting lyophilized protecting agent.
Background technology
Freeze Drying Technique is biological a kind of important application technology, is usually used in the research and production of biological product, to protect
Hold the biological characteristics of bacterium, seed culture of viruses, stabilate raw material and the biological activity of semi-finished product, extend the effect duration of finished product.Lyophilizing
Protective agent is to give effective ingredient composition of storage stage protection medicine effect after freezing dry process and lyophilizing, is alive
Key technology in vaccine or antibody producing, for vaccine or the preservation of antibody, transports and use has a very big impact.
Veterinary biologics typically requires cryopreservation, is only capable of preserving about 6 months under the conditions of 4 DEG C, current in conjunction with China
Aquaculture level, quite a few raiser does not possess the primary condition of cryopreservation biological product, the most such as
Fruit can not be finished in time, easily causes the inefficacy of biological product, brings economic loss to raiser.At present, China's life for animals
The freeze drying protectant of Tetramune is mainly composed of the gelatin of the sucrose of skim milk, 5%～10%, 1.5%～3% of 5%, group
Side is simple, and defencive function is poor.On market, existing freeze drying protectant is 18 months to the longest protection period of yolk antibody,
Have impact on the performance of yolk antibody effect, and the freeze drying process of infectious bursal disease yolk antibody to a certain extent
Yolk antibody is caused injury in various degree by the ice crystal formed, and directly affects the activity of antibody in dried product, and then
Have influence on the quality of product.
Summary of the invention
It is an object of the invention to provide a kind of infectious bursal disease yolk antibody heat-resisting lyophilized protecting agent, extend yolk antibody
Pot-life, effectively keep the curative effect of yolk antibody.
Second object of the present invention is to provide the preparation of a kind of infectious bursal disease yolk antibody heat-resisting lyophilized protecting agent
Method.
In order to realize object above, the technical solution adopted in the present invention is: a kind of infectious bursal disease yolk antibody is resistance to
Hot freeze drying protectant, consists of the following components in 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
For water for injection.
The preparation method of a kind of above-mentioned infectious bursal disease yolk antibody heat-resisting lyophilized protecting agent, comprises the following steps:
1) take 1/3rd of the total consumption of water for injection, add oligomeric xylose, sorbitol, mix homogeneously, obtain mixture A；
2) take 1/3rd of the total consumption of water for injection, add sodium carboxymethyl cellulose, mix homogeneously, obtain mixture B；
3) take remaining water for injection, add sodium glutamate, sodium selenite, glycerol, mix homogeneously, obtain mixture C；
4) it is mixed and stirred for uniformly, obtaining final product by mixture A, mixture B, mixture C.
The infectious bursal disease yolk antibody heat-resisting lyophilized protecting agent of the present invention, during use, by heat-resisting lyophilized protecting agent with
Infectious bursal disease yolk antibody mixes according to the ratio that volume ratio is 3～5:1, carries out freezing dry after subpackage
Dry.
The 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 compound with glycerol, make infectious bursal disease yolk antibody at 2～8 DEG C of bars
Can preserve 36 months under part, the profile of yolk antibody, activity, and antibody effect can be kept during freeze-drying process and preservation
Valency does not has significant change, and before and after lyophilizing, yolk antibody loss rate is low, and freeze-drying prods has good ageing-resistant performance, slow down
The speed of yolk antibody activity decrease, avoids 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, by following components in percentage by weight group
Become: oligomeric xylose 8%, sodium carboxymethyl cellulose 2%, sodium glutamate 2%, sorbitol 3%, sodium selenite 0.5%, sweet
Oil 2%, surplus is water for injection.
The preparation method of a kind of above-mentioned infectious bursal disease yolk antibody heat-resisting lyophilized protecting agent, comprises the following steps:
1) take 1/3rd of the total consumption of water for injection, add oligomeric xylose, sorbitol, mix homogeneously, obtain mixture A；
2) take 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) take remaining water for injection, add sodium glutamate, sodium selenite, glycerol, mix homogeneously, obtain mixture C；
4) it is mixed and stirred for uniformly, obtaining heat-resisting lyophilized protecting agent by mixture A, mixture B, mixture C.
By the present embodiment gained heat-resisting lyophilized protecting agent with avian infectious bursal disease yolk antibody according to the ratio that volume ratio is 3:1
Example mixes, and carries out lyophilization after subpackage, and rear sealing, packaging products obtained therefrom can preserve 30 under the conditions of 2～8 DEG C
Individual month.
Embodiment 2
The infectious bursal disease yolk antibody heat-resisting lyophilized protecting agent of the present embodiment, by following components in percentage by weight group
Become: oligomeric xylose 9%, sodium carboxymethyl cellulose 2.5%, sodium glutamate 3%, sorbitol 4%, sodium selenite 0.6%, sweet
Oil 8%, surplus is water for injection.
The preparation method of a kind of above-mentioned infectious bursal disease yolk antibody heat-resisting lyophilized protecting agent, comprises the following steps:
1) take 1/3rd of the total consumption of water for injection, add oligomeric xylose, sorbitol, mix homogeneously, obtain mixture A；
2) take 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) take remaining water for injection, add sodium glutamate, sodium selenite, glycerol, mix homogeneously, obtain mixture C；
4) it is mixed and stirred for uniformly, obtaining heat-resisting lyophilized protecting agent by mixture A, mixture B, mixture C.
By the present embodiment gained heat-resisting lyophilized protecting agent with avian infectious bursal disease yolk antibody according to the ratio that volume ratio is 4:1
Example mixes, and carries out lyophilization after subpackage, and rear sealing, packaging products obtained therefrom can preserve 36 under the conditions of 2～8 DEG C
Individual month.
Embodiment 3
The infectious bursal disease yolk antibody heat-resisting lyophilized protecting agent of the present embodiment, by following components in percentage by weight group
Become: oligomeric xylose 10%, sodium carboxymethyl cellulose 3%, sodium glutamate 4%, sorbitol 5%, sodium selenite 0.8%, sweet
Oil 4%, surplus is water for injection.
The preparation method of a kind of above-mentioned infectious bursal disease yolk antibody heat-resisting lyophilized protecting agent, comprises the following steps:
1) take 1/3rd of the total consumption of water for injection, add oligomeric xylose, sorbitol, mix homogeneously, obtain mixture A；
2) take 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) take remaining water for injection, add sodium glutamate, sodium selenite, glycerol, mix homogeneously, obtain mixture C；
4) it is mixed and stirred for uniformly, obtaining heat-resisting lyophilized protecting agent by mixture A, mixture B, mixture C.
By the present embodiment gained heat-resisting lyophilized protecting agent with avian infectious bursal disease yolk antibody according to the ratio that volume ratio is 5:1
Example mixes, and carries out lyophilization after subpackage, and rear sealing, packaging products obtained therefrom can preserve 30 under the conditions of 2～8 DEG C
Individual month.
Experimental example
This experimental example uses the freeze drying protectant preparation infectious bursal disease yolk antibody of embodiment 1～3 and carries out lyophilizing
It is dried, is respectively placed in 2～8 DEG C, preserves under the conditions of 37 DEG C, detect yolk antibody titer situation of change in different time.
Experimental technique: by be saved in 2～8 DEG C, infectious bursal disease yolk antibody under the conditions of 37 DEG C periodically take out, press
Label indicates plumage part, by antibody normal saline dilution to 1 plumage part ml.8.0% sodium chloride solution 100ml adds
1g high-quality agar powder, after heating and melting, adds final concentration of 0.01% thimerosal anticorrosion, casts gel slab, thereon by six
Dihedral punches, and aperture is 5mm, and pitch-row is 3mm.Test after antibody is made 2 times of serial dilutions, antigen and antibody
Dosage is with Kong Manwei degree (about 25ul)；Then agar plate is put in wet box, hatch 24～28 hours for 37 DEG C.Work as yin, yang
Property serum control when setting up, the antibody maximum dilution multiple forming white precipitate line with standard antigen is the effect of yolk antibody
Valency.
In this experiment, matched group 1 is that the chicken of the common freeze drying protectant preparation using conventional 5% sucrose skimmed milk preparation is infected
Property bursal disease yolk antibody；Matched group 2 is that the infectious bursal disease that Shandong company produces refines yolk antibody；Real
Testing group 1,2,3 is the infectious bursal disease utilizing the embodiment of the present application 1,2,3 gained heat-resisting lyophilized protecting agent to prepare
Yolk antibody.
Experimental result is as shown in table 1,2.
Table 1 yolk antibody preserves 6,12,18,24,30,36 months titration results under the conditions of 2～8 DEG C
| Group | Basis titer | 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 the conditions of 37 DEG C
| Group | Basis titer | 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 |
From table 1,2 it can be seen that use the avian infectious method of the embodiment of the present application 1～3 gained heat-resisting lyophilized protecting agent preparation
Family name's capsule disease yolk antibody, at different temperatures than the infectious bursal disease yolk antibody prepared with common freeze drying protectant with
And commercially available infectious bursal disease refines the good stability of yolk antibody titer, the holding time of antibody titer extends.Examination
Test result explanation, use the infectious bursal disease yolk antibody of the heat-resisting lyophilized protecting agent preparation of the application to have good
Thermostability, antibody titer has more preferable stability.
Claims (2)
1. an infectious bursal disease yolk antibody heat-resisting lyophilized protecting agent, it is characterised in that: by following weight percent
The component composition of ratio: oligomeric xylose 8%～10%, sodium carboxymethyl cellulose 2%～3%, sodium glutamate 2%～4%, Pyrusussuriensis
Alcohol 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,
It is characterized in that: comprise the following steps:
1) take 1/3rd of the total consumption of water for injection, add oligomeric xylose, sorbitol, mix homogeneously, obtain mixture A；
2) take 1/3rd of the total consumption of water for injection, add sodium carboxymethyl cellulose, mix homogeneously, obtain mixture B；
3) take remaining water for injection, add sodium glutamate, sodium selenite, glycerol, mix homogeneously, obtain mixture C；
4) it is mixed and stirred for uniformly, obtaining final product by mixture A, mixture B, mixture C.
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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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| 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 |
Non-Patent Citations (1)
| Title |
|---|
| 耐热冻干保护剂及其在兽用冻干活疫苗中的应用;杨泽林等;《四川畜牧兽医》;20031231;第30卷(第12期);33-34 * |
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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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