CN1426816A - Veterinarian virus kind biological product heat resisting freeze drying protective agent and its preparation technique - Google Patents
Veterinarian virus kind biological product heat resisting freeze drying protective agent and its preparation technique Download PDFInfo
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Abstract
A freeze-dried high-temp. resistant protecting agent for virus-type biologic products for veterinary medicine is prepared from several components through proportionally mixing. It features that each of its components is sterilized separately. For the components which can be sterilized under high temp., they are dissolved in distilled water according to the proportion of formulation and are sterilized at 116 deg.C for 30-40 min; and for those which do not resist the high temp., they are also dissolved in distilled water according the proportion of formulation and are filtered with 0.22 micron pore diameter filter membrane to remove bacteria; and then the two parts are mixed to obtain the freeze-dried high-temp. resistant, protecting agent. The mentioned two parts are added to the virus antigen liquid according to a proportion of 1:1 and after packaging, they are freeze-dried. The product can be stored at 2-8 deg.C for 24 months.
Description
Affiliated technical field
The present invention relates to the Freeze Drying Technique of biological product, preservation adjuvant and manufacture method thereof.
Background technology
China's veterinary biological product protective agent commonly used at present is mainly 5% sucrose skim milk and 5-10% sucrose, the 1.5-3% gelatin, and goods require cryopreservation (below 15 ℃), and the storage life of 2-8 ℃ of condition only 3-9 month must use up at normal temperatures as early as possible.This is just for the preservation of veterinary biological product, transportation, use etc. bring many difficulties and inconvenience, simultaneously owing to must cryopreservation causing energy resource consumption and storage cost big.
At present, the flourishing in the world biological freeze-dried products of national veterinary almost all is that 2-8 ℃ of preservation is more than 24 months.But the not play-by-play of square method.Chinese patent CN1176827A " a kind of manufacture method of freeze-dried live vaccine for animal use " proposes to adopt with polyvinylpyrrolidone, sodium carboxymethyl cellulose; tamarind gum; sodium glutamate; trehaloses etc. are the freeze drying protectant that the formulated of main matrix freeze drying protectant becomes; the method that freeze-drying curve that employing adapts with it and newcastle disease vaccine virus antigen are made the newcastle disease freeze-dried live vaccine, but this Seedling long preservation under 2-8 ℃ of condition.
Summary of the invention
Veterinarian virus kind biological product heat resisting freeze drying protective agent and preparation technology, comprise that various heat-resisting lyophilized protecting agents with Veterinarian virus kind biological product are respectively by component, mixed, it is characterized in that each protectant active princlple is handled degerming respectively:, sterilized 30-40 minute for 116 ℃ to can autoclavedly being dissolved in the distilled water in the preparation ratio; The component of non-refractory also is dissolved in the distilled water with aperture 0.22 μ m membrane filtration degerming in the preparation ratio; After finishing degerming respectively two kinds of compositions are mixed, be the lyophilizing heat resisting protective, add in the virus antigen liquid in 1: 1 ratio during use, carry out lyophilization after the packing.
Preparation technology of the present invention is the characteristics according to different virus, create the combination of different component, different proportion, technological requirement is formulated rational freeze-drying curve routinely, to guarantee the biological activity of goods, various preparation of the present invention is preserved more than 24 months under 2-8 ℃ of condition, all can reach every index of regulation in " People's Republic of China's veterinary biological product quality standard ".
The specific embodiment
Veterinarian virus kind biological product heat resisting freeze drying protective agent, protectant component comprises gelatin commonly used, sucrose, defatted milk powder, dextrin, it is characterized in that the component proportioning of various heat-resisting lyophilized protecting agents is: the weak malicious freeze dried vaccine of (1) newcastle disease: gelatin 1.5-2%, sucrose 5-6%, dextrin 5-10%, PVP (K90) 2-5%, sorbitol 2%, sodium glutamate 1-2%; (2) the weak poison of infectious bursal disease (B87) freeze dried vaccine: defatted milk powder 5%, dextrin 5%, beef peptone 1-3%, sucrose 5-6%, sorbitol 2%, sodium glutamate 1-3%, Vc0.1%; (3) infectious bronchitis of chicken (H120, H52, Kidney infectious bronchitis W93) freeze dried vaccine: gelatin 1.5-2%, bovine serum albumin 1-2%, sucrose 5-6%, PVP (K90) 5%, casein peptone 2%, sorbitol 1-3%, sodium glutamate 1-2%, Vc0.1-0.3%; (4) newcastle disease, infectious bronchitis of chicken (H120, H52) bigeminy freeze dried vaccine: gelatin 2%, dextrin 6%, bovine serum albumin 1-2%, sucrose 5-6%, PVP (K90) 3%, casein peptone 1-2%, sorbitol 2%, sodium glutamate 1%, Vc0.2-0.3%; (5) infectious laryngotracheitis of chicken freeze dried vaccine: gelatin 2%, dextrin 5%, lactose 5-6%, sorbitol 2%, arginase 12 %, sodium glutamate 1%, Vc0.1%; (6) swine fever bovine testicle cell Seedling: gelatin 3%, lactose 5-6%, beef peptone 2%, PVP (K90) 3%, sorbitol 2%, sodium glutamate 2%, Vc0.1%; (7) fowlpox freeze-dried vaccine: gelatin 3%, EDTA2%, lactose 5-6%, beef peptone 2%, bovine serum albumin 1%, PVP (K90) 5%, sorbitol 2%, sodium glutamate 1%, Vc0.1%; (8) marek isease turkey herpes virus freeze-dried vaccine: gelatin 1.5%, EDTA2%, sucrose 7.6%, beef peptone 1%, bovine serum albumin 1%.Be the dry component in above-mentioned each preparation, percentage ratio residue part is distilled water.
The preparation technology of Veterinarian virus kind biological product heat resisting freeze drying protective agent, comprise various heat-resisting lyophilized protecting agents respectively by component, mixed, it is characterized in that each protectant active princlple is handled degerming respectively: to can be autoclaved being dissolved in the distilled water in ratio separately, 116 ℃ of sterilizations 30-40 minute; The component of non-refractory also is dissolved in the distilled water in proportion with aperture 0.22 μ m membrane filtration degerming; After finishing degerming respectively two kinds of compositions are mixed, be the lyophilizing heat resisting protective, add in the virus antigen liquid in 1: 1 ratio during use, carry out lyophilization after the packing.But protectant active princlple is divided into autoclaving and thermo-labile filtering two classes by degerming technology: (1) but the weak malicious freeze dried vaccine autoclaving component of newcastle disease be: gelatin, sucrose, dextrin, PVP (K90); The component of thermo-labile filtration sterilization is: sorbitol, sodium glutamate; (2) the weak poison of infectious bursal disease (B87) but freeze dried vaccine autoclaving component be: defatted milk powder, dextrin, beef peptone, sucrose; The component of thermo-labile filtration sterilization is: sorbitol, sodium glutamate, Vc; (3) infectious bronchitis of chicken (H120, H52, Kidney infectious bronchitis W93) but freeze dried vaccine autoclaving component be: gelatin, sucrose, PVP (K90), casein peptone; The component of thermo-labile filtration sterilization is: bovine serum albumin, sorbitol, sodium glutamate, Vc; (4) newcastle disease, infectious bronchitis of chicken (H120, H52) but bigeminy freeze dried vaccine autoclaving component be: gelatin, dextrin, sucrose, PVP (K90), casein peptone; The component of thermo-labile filtration sterilization is: bovine serum albumin, sorbitol, sodium glutamate, Vc; (5) but infectious laryngotracheitis of chicken freeze dried vaccine autoclaving component be: gelatin, dextrin, lactose; The component of thermo-labile filtration sterilization is: sorbitol, arginine, sodium glutamate, Vc; (6) but swine fever bovine testicle cell Seedling autoclaving component be: gelatin, lactose, beef peptone, PVP (K90); The component of thermo-labile filtration sterilization is: sorbitol, sodium glutamate, Vc; (7) but fowlpox freeze-dried vaccine autoclaving component be: gelatin, EDTA, lactose, beef peptone, PVP (K90); The component of thermo-labile filtration sterilization is: bovine serum albumin, sorbitol, sodium glutamate, Vc; (8) but marek isease turkey herpes virus freeze-dried vaccine autoclaving component be: gelatin, EDTA, sucrose, beef peptone; The component of thermo-labile filtration sterilization is: phosphorus potassium dihydrogen, dipotassium hydrogen phosphate, sorbitol, sodium glutamate, bovine serum albumin.
Technical parameter and detection method
1. the physical behavior of product, steriling test, mycoplasma check, exogenous virus check, specificity check, safety verification, residual moisture mensuration and vacuum are measured by the method and the standard of " People's Republic of China's veterinary biological product rules " and are undertaken.
2. lyophilization cycle is no more than 48h.
3. protectant toxicity test
Bird vaccine carries out each 20 of collunarium, eye dripping, drinking-water and subcutaneous injections by containing 10 times of amounts protecting dosage in the middle of the product respectively to 7 age in days healthy chicks, observes 20d, and chickling is all strong to live, and does not have any untoward reaction.
4. the viral level of each product is measured and standard
1) the weak malicious freeze-dried live vaccine of newcastle disease
(1) assay method:
A. make 10 times of serial dilutions by the real tissue mass that contains with sterile saline, get 3 suitable dilution factors, inoculate 5 of 10 age in days SPF Embryo Gallus domesticus in the allantoic cavity respectively, every embryo 0.1mL puts 37 ℃ and continues to hatch, the dead in the past Embryo Gallus domesticus of 48h discards to be disregarded, at the Embryo Gallus domesticus of 48-120h death, take out results Embryo Gallus domesticus liquid at any time, with same dilution Embryo Gallus domesticus liquid mixed in equal amounts, measure the red cell agglutination valency respectively.To 120h, get all embryos of living, gather in the crops Embryo Gallus domesticus liquid one by one, measure the red cell agglutination valency respectively, agglutination titer 〉=160 are judged to infection, calculate EID
50
B. press label and indicate plumage part, vaccine is diluted to 1 plumage part/0.1mL with sterile saline, remake 10 times of serial dilutions, other method is the same, calculates every plumage part EID at last
50Content.
(2) reach standard
A. after the lyophilizing viral loss rate less than 0.4 titre.
B. every plumage part viral level 〉=10 after the lyophilizing
6.7EID
50
C. the product after the lyophilizing is put 37 ℃ and is preserved 10d and 30d and carry out viral level and measure, rate of descent the former within 1 titre, the latter is within 2 titres.
D. the product after the lyophilizing is put 25 ℃ of preservation 30d, and the viral level rate of descent is within 0.6 titre.
E. product is put 2-8 ℃ of preservation 24 months, every plumage part viral level 〉=10
6.17EID
50Chicken
2) the weak poison of infectious bursal disease (B
87) the tissue freeze-dried vaccine
(1) assay method: vaccine is made 10 times of serial dilutions with sterile saline, and with allantocherion approach inoculation 10-12 age in days SPF Embryo Gallus domesticus, every embryo 0.2mL observes 168h, calculates ELD
50
3. protectant toxicity test
Bird vaccine carries out each 20 of collunarium, eye dripping, drinking-water and subcutaneous injections by containing 10 times of amounts protecting dosage in the middle of the product respectively to 7 age in days healthy chicks, observes 20d, and chickling is all strong to live, and does not have any untoward reaction.
4. the viral level of each product is measured and standard
1) the weak malicious freeze-dried live vaccine of newcastle disease
(1) assay method:
A. make 10 times of serial dilutions by the real tissue mass that contains with sterile saline, get 3 suitable dilution factors, inoculate 5 of 10 age in days SPF Embryo Gallus domesticus in the allantoic cavity respectively, every embryo 0.1mL puts 37 ℃ and continues to hatch, and dead Embryo Gallus domesticus discards and disregards before the 48h, 48
-The Embryo Gallus domesticus of 120h death takes out at any time, and results Embryo Gallus domesticus liquid with same dilution Embryo Gallus domesticus liquid mixed in equal amounts, is measured the red cell agglutination valency respectively.To 120h, get all embryos of living, gather in the crops Embryo Gallus domesticus liquid one by one, measure the red cell agglutination valency respectively, agglutination titer 〉=160 are judged to infection, calculate EID
50
B. press label and indicate plumage part, vaccine is diluted to 1 plumage part/0.1mL with sterile saline, remake 10 times of serial dilutions, other method is the same, calculates every plumage part EID at last
50Content.
(2) reach standard
A. after the lyophilizing viral loss rate less than 0.4 titre.
B. every plumage part viral level 〉=10 after the lyophilizing
6.7EID
50
C. the product after the lyophilizing is put 37 ℃ and is preserved 10d and 30d and carry out viral level and measure, rate of descent the former within 1 titre, the latter is within 2 titres.
D. the product after the lyophilizing is put 25 ℃ of preservation 30d, and the viral level rate of descent is within 0.6 titre.
E. product is put 2-8 ℃ of preservation 24 months, every plumage part viral level 〉=10
6.17EID
50Chicken
2) the weak poison of infectious bursal disease (B
87) the tissue freeze-dried vaccine
(1) assay method: vaccine is made 10 times of serial dilutions with sterile saline, and with allantocherion approach inoculation 10-12 age in days SPF Embryo Gallus domesticus, every embryo 0.2mL observes 168h, calculates ELD
50
(2) reach standard
A. after the lyophilizing viral loss rate less than 0.4 titre.
B. every plumage part viral level 〉=10 after the lyophilizing
3.7ELD
50
C. the product after the lyophilizing is put 37 ℃ and is preserved 10d and 30d and carry out viral level and measure, and the former rate of descent is within 0.2 titre, and the latter is within 0.4 titre.
D. product is put 25 ℃ to preserve 30d viral level rates of descent is within 0.2 titre.
E. product was preserved every plumage part viral level 〉=10 24 months at 2-8 ℃
3.37ELD
50
3) infectious bronchitis of chicken (H
120, H
52, Kidney infectious bronchitis W
93) freeze dried vaccine
(1) assay method:
A. make 10 times of serial dilutions with normal saline by the real tissue mass that contains, get 3 dilution factors and respectively inoculate 5 of instar chicken embryos on the 10th, inoculation 0.1mL in every embryo allantoic cavity, putting 37 ℃ continues to hatch, the dead in the past Embryo Gallus domesticus of 24h discards to be disregarded, specificity lesion persons' such as in the embryo alive of still surviving according to inoculation back 24-144h dead germ and 144h, its fetus has dehydration, rolls up, dysplasia summation is calculated its EID
50
B. press label and indicate plumage part, vaccine is diluted to 1 plumage part/0.1mL with normal saline, do 10 times of serial dilutions again, following method is the same.
(2) reach standard
A. lyophilizing virus loss rate is again within 0.4 titre.
B. H after the lyophilizing
120, H
52Every plumage part viral level 〉=10
4.5EID
50Kidney infectious bronchitis W
93Every plumage part viral level 〉=10
5.5EID
50
C. product is put 37 ℃ of conditions preservation 10d and 30d, and the viral level rate of descent is respectively within 1 and 2 titre.
D. product is put 25 ℃ of conditions preservation 30d, and the viral level rate of descent is no more than 1 titre.
E. product is put 2-8 ℃ of preservation 24 months, H
120, H
52Every plumage part viral level 〉=10
3.5EID
50, Kidney infectious bronchitis W
93Every plumage part viral level 〉=10
4.67EID
50
4. newcastle disease, infectious bronchitis of chicken (H
120, H
52) bigeminal live vaccine
(1) assay method:
A. vaccine is diluted to 1 plumage part/0.5mL with normal saline, is respectively charged into 2 in vitro, every pipe 1mL.The 1st pipe adds the newcastle disease antiserum of equivalent, and the 2nd pipe adds the infectious bronchitis of chicken antiserum of equivalent.At room temperature and 1h, this moment, viral level was 0.1 plumage part/0.1mL, promptly 10
-1The 1st pipe continues 10 times of serial dilutions.Measure the infectious bronchitis of chicken part.The 2nd pipe continues 10 times of serial dilutions, measures the newcastle disease part, and its method is with infectious bronchitis of chicken and newcastle disease freeze-dried vaccine.
B. vaccine is contained tissue mass and is diluted to 500 times by real with normal saline, be respectively charged in 2 test tubes, every pipe 1mL, the 1st pipe adds the newcastle disease antiserum of equivalent, and the 2nd pipe adds equivalent infectious bronchitis of chicken antiserum.Be 10 this moment
-3, remaking 10 times of serial dilutions at room temperature with behind the 1h, other method is the same.
(2) reach standard
Respectively with malicious freeze-dried vaccine a little less than the newcastle disease and infectious bronchitis of chicken freeze-dried vaccine effect.
5 infectious laryngotracheitis of chicken freeze dried vaccines
(1) method of inspection:
A. press label and indicate plumage part, vaccine is diluted to 1 plumage part 0.2mL with sterile saline, continue again to make 10 times of serial dilutions, get 3 suitable dilution factors, each allantocherion inoculation 10
-Instar chicken embryo was 5 on 11st, every embryo 0.2mL, and 5d is cultivated in the inoculation back, dissects, and observes pathological changes, and chick chorioallantoic membrane obviously thickens, and the canescence scab is arranged, and is judged to infection, calculates EID
50
B. by the real tissue mass 10 times of serial dilutions of sterile saline that contain, other method is the same.
(2) reach standard
A. lyophilizing virus loss rate is within 0.4 titre.
B. every plumage part viral level 〉=10 after the lyophilizing
3.7EID
50
C. product is put 37 ℃ of condition 10d and 30d, and viral loss rate is respectively within 1 and 1.5 titre.
D. product is put 25 ℃ of condition 30d, and viral loss rate is within 1 titre.
E. product 2-8 ℃ of condition preserved every plumage part viral level 〉=10 24 months
2.83EID
50
6 swine fever bovine testicle cell freeze dried vaccines
(1) detection method:
Imitate inspection with rabbit: press the dated head part of label, dilute suitable multiple with 750 times of every part vaccine dilutions or by the real antigen amount that contains with normal saline, 2 of ear vein injection rabbit, every 1mL changes to determine tiring of vaccine according to rabbit body temperature.
(2) reach standard
A. after the lyophilizing vaccine reach 7.5 ten thousand times qualified.
B. product is preserved 10d and 30d under 37 ℃ of conditions, and it is qualified that the effectiveness of product reaches 50,000 times and 40,000 times.
C. product is preserved 30d under 25 ℃ of conditions, and 50,000 times qualified.
D. product was preserved 24 months under 2-8 ℃ of condition, and the effectiveness of product is all more than 50,000 times.
7 fowlpox freeze-dried live vaccines
(1) detection method:
A. press label and indicate plumage part, with the normal saline dilution, 10 of allantocherion inoculation 11-12 day instar chicken embryos, every embryo 0.2mL (containing 1/100 using dosage).Behind the 96-120h, the chick chorioallantoic membrane edema thickens or the pox speckle occurs to be judged.
B. make 10 times of serial dilutions by the real antigen amount that contains with normal saline, get 3 suitable dilution factors, the every embryo 0.2mL of inoculation 11-12 age in days chick chorioallantoic membrane, thickening or the pox speckle occurs according to the chick chorioallantoic membrane edema behind the 96-120h is to infect to judge EID
50
(2) reach standard
A. lyophilizing virus loss rate is 0.4 below the titre.
B. EID after the lyophilizing
50All 〉=10
6.1EID
50/ 0.2mL.
C. product is preserved 10d and 30d under 37 ℃ of conditions, and viral loss rate is respectively within 0.8 and 1.2 titre.
D. product is preserved 30d under 25 ℃ of conditions, and viral loss rate is within 0.6 titre.
E. product was preserved 24 months 2-8 ℃ of condition, measured 10 embryos by " A " method and all obviously infected.
8 marek isease turkey herpes virus freeze dried vaccines
(1) detection method:
With " veterinary biologics rules ", press the method for plaque forming unit (PFU) and measure.
(2) reach standard
A. lyophilizing loss, the plaque slip is below 45%.
B. product is preserved 10d under 37 ℃ of conditions, and the plaque slip is below 50%.
C. product is preserved 20d under 25 ℃ of conditions, and the plaque slip is below 20%.
D. after the lyophilizing every plumage part viral level all greater than 5000PFU.
E. product was preserved 24 months 2-8 ℃ of condition, and every plumage part viral level is all greater than 3000PFU.
Claims (3)
1, a kind of Veterinarian virus kind is held the Tetramune heat-resisting lyophilized protecting agent, protectant component comprises sucrose commonly used, defatted milk powder, gelatin, dextrin, it is characterized in that various protectant component proportionings are: the weak malicious freeze dried vaccine of (1) newcastle disease: gelatin 1.5-2%, sucrose 5-6%, dextrin 5-10%, PVP (K90) 2-5%, sorbitol 2%, sodium glutamate 1-2%; (2) the weak poison of infectious bursal disease (B87) freeze dried vaccine: defatted milk powder 5%, dextrin 5%, beef peptone 1-3%, sucrose 5-6%, sorbitol 2%, sodium glutamate 1-3%, Vc0.1%; (3) infectious bronchitis of chicken (H120, H52, Kidney infectious bronchitis W93) freeze dried vaccine: gelatin 1.5-2%, bovine serum albumin 1-2%, sucrose 5-6%, PVP (K90) 5%, casein peptone 2%, sorbitol 1-3%, sodium glutamate 1-2%, Vc0.1-0.3%; (4) newcastle disease, infectious bronchitis of chicken (H120, H52) bigeminy freeze dried vaccine: gelatin 2%, dextrin 6%, bovine serum albumin 1-2%, sucrose 5-6%, PVP (K90) 3%, casein peptone 1-2%, sorbitol 2%, sodium glutamate 1%, Vc0.2-0.3%; (5) infectious laryngotracheitis of chicken freeze dried vaccine: gelatin 2%, dextrin 5%, lactose 5-6%, sorbitol 2%, arginase 12 %, sodium glutamate 1%, Vc0.1%; (6) swine fever bovine testicle cell Seedling: gelatin 3%, lactose 5-6%, beef peptone 2%, PVP (K90) 3%, sorbitol 2%, sodium glutamate 2%, Vc0.1%; (7) fowlpox freeze-dried vaccine: gelatin 3%, EDTA2%, lactose 5-6%, beef peptone 2%, bovine serum albumin 1%, PVP (K90) 5%, sorbitol 2%, sodium glutamate 1%, Vc0.1%; (8) marek isease turkey herpes virus freeze-dried vaccine: gelatin 1.5%, EDTA2%, sucrose 7.6%, beef peptone 1%, bovine serum albumin 1%, phosphorus potassium dihydrogen 0.52%, dipotassium hydrogen phosphate 1.64%, sorbitol 1%, sodium glutamate 1%; Be the dry component in above-mentioned each preparation, percentage ratio residue part is distilled water.
2, a kind of preparation technology of Veterinarian virus kind biological product heat resisting freeze drying protective agent, comprise right 1 described various protective agents respectively by component, mixed, it is characterized in that respectively each protectant active princlple being handled degerming respectively:, sterilized 30-40 minute for 116 ℃ to can autoclavedly being dissolved in the distilled water in the preparation ratio; The component of non-refractory also is dissolved in the distilled water with aperture 0.22 μ m membrane filtration degerming in the preparation ratio; After finishing degerming respectively two kinds of compositions are mixed, be the lyophilizing heat resisting protective, add in the virus antigen liquid in 1: 1 ratio during use, carry out lyophilization after the packing.
3, the preparation technology of Veterinarian virus kind biological product heat resisting freeze drying protective agent according to claim 2, but it is characterized in that various protectant active princlples are divided into autoclaving and thermo-labile filtering two classes by degerming technology: (1) but the weak malicious freeze dried vaccine autoclaving component of newcastle disease be: gelatin, sucrose, dextrin, PVP (K90); The component of thermo-labile filtration sterilization is: sorbitol, sodium glutamate; (2) the weak poison of infectious bursal disease (B87) but freeze dried vaccine autoclaving component be: defatted milk powder, dextrin, beef peptone, sucrose; The component of thermo-labile filtration sterilization is: sorbitol, sodium glutamate, Vc; (3) infectious bronchitis of chicken (H120, H52, Kidney infectious bronchitis W93) but freeze dried vaccine autoclaving component be: gelatin, sucrose, PVP (K90), casein peptone; The component of thermo-labile filtration sterilization is: bovine serum albumin, sorbitol, sodium glutamate, Vc; (4) newcastle disease, infectious bronchitis of chicken (H120, H52) but bigeminy freeze dried vaccine autoclaving component be: gelatin, dextrin, sucrose, PVP (K90), casein peptone; The component of thermo-labile filtration sterilization is: bovine serum albumin, sorbitol, sodium glutamate, Vc; (5) but infectious laryngotracheitis of chicken freeze dried vaccine autoclaving component be: gelatin, dextrin, lactose; The component of thermo-labile filtration sterilization is: sorbitol, arginine, sodium glutamate, Vc; (6) but swine fever bovine testicle cell Seedling autoclaving component be: gelatin, lactose, beef peptone, PVP (K90); The component of thermo-labile filtration sterilization is: sorbitol, sodium glutamate, Vc; (7) but fowlpox freeze-dried vaccine autoclaving component be: gelatin, EDTA, lactose, beef peptone, PVP (K90); The component of thermo-labile filtration sterilization is: bovine serum albumin, sorbitol, sodium glutamate, Vc; (8) but marek isease turkey herpes virus freeze-dried vaccine autoclaving component be: gelatin, EDTA, sucrose, beef peptone; The component of thermo-labile filtration sterilization is: phosphorus potassium dihydrogen, dipotassium hydrogen phosphate, sorbitol, sodium glutamate, bovine serum albumin.
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CNB011388714A CN1168503C (en) | 2001-12-21 | 2001-12-21 | Veterinarian virus kind biological product heat resisting freeze drying protective agent and its preparation technique |
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CNB011388714A CN1168503C (en) | 2001-12-21 | 2001-12-21 | Veterinarian virus kind biological product heat resisting freeze drying protective agent and its preparation technique |
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CNB2004100295841A Division CN1222320C (en) | 2001-12-21 | 2001-12-21 | Heat-resistant lyophilized protectant of lyophilized vaccine for chicken infectious laryngotracheitis and preparing process thereof |
CNB2004100295837A Division CN1223375C (en) | 2001-12-21 | 2001-12-21 | Heat resisting lyophilized protectant for chicken infectious bronchitis (H120, H52, Kidney infectious bronchitis W93) lyophilized vaccine and preparing process thereof |
CN 200410029587 Division CN1222314C (en) | 2001-12-21 | 2001-12-21 | Heat resisting lyophilized protectant for chicken Marek's disense turkey herpes virus lyophilized vaccine and preparing process thereof |
CNB2004100295822A Division CN1233418C (en) | 2001-12-21 | 2001-12-21 | Heat-resisting lyophilized protectant for chicken newcastle disease, chicken infectious bronchitis (H120, H52) dual lyophilized vaccine and preparing process thereof |
CN 200410029581 Division CN1251762C (en) | 2001-12-21 | 2001-12-21 | Heat resisting lyophilized protectant for chicken infections Fabianite bursa hypotpxicity (B87) lyophilized vaccine and preparing process thereof |
CNB2004100295860A Division CN1253209C (en) | 2001-12-21 | 2001-12-21 | Heat-resistant lyophilized protectant of lyophilized vaccine for henpox and preparing process thereof |
CN 200410029585 Division CN1211122C (en) | 2001-12-21 | 2001-12-21 | Heat-resisting lyophilized protectant for swine fever lapinized hyoptoxicity calf testicular cell vaccine and preparing process |
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CN1168503C CN1168503C (en) | 2004-09-29 |
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