CN102240400A - Method for producing rabies vaccine by applying bioreactor and sheet carrier - Google Patents
Method for producing rabies vaccine by applying bioreactor and sheet carrier Download PDFInfo
- Publication number
- CN102240400A CN102240400A CN201110137119XA CN201110137119A CN102240400A CN 102240400 A CN102240400 A CN 102240400A CN 201110137119X A CN201110137119X A CN 201110137119XA CN 201110137119 A CN201110137119 A CN 201110137119A CN 102240400 A CN102240400 A CN 102240400A
- Authority
- CN
- China
- Prior art keywords
- cell
- virus
- vaccine
- rabies
- rabies vaccine
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
本发明涉及一种应用生物反应器及片状载体生产狂犬疫苗的方法，属于生物制品技术领域。在具有固定床篮式搅拌系统的Celligen310型14L生物反应器中以台湾赛宇細胞科技股份有限公司的BioNOCII?cellculturedisks为载体培养细胞，通过特定的上罐细胞密度、所加病毒和生物反应器设定参数制备高滴度的狂犬病毒，生产狂犬疫苗。该方法具有培养的细胞密度高、病毒滴度和产量高以及生产成本低的优点，此反应器通过加入BioNOCII?cellculturedisks增加可供细胞贴壁生长的表面积，再通过创造良好的培养条件使细胞培养密度达到5×108个/ml，病毒滴度相应的大幅度成对数增加，最高可达1×109FFU/ml，大幅度地降低了生产成本，适用于狂犬疫苗的大批量生产。The invention relates to a method for producing rabies vaccine using a bioreactor and a sheet carrier, belonging to the technical field of biological products. In the Celligen310 14L bioreactor with a fixed bed basket stirring system, the cells were cultured with the BioNOCII? cellculturedisks of Taiwan Saiyu Cell Technology Co., Ltd. as the carrier. Set parameters to prepare high-titer rabies virus and produce rabies vaccine. This method has the advantages of high cultured cell density, high virus titer and yield, and low production cost. The reactor increases the surface area for cell growth by adding BioNOCII? When the density reaches 5×10 8 cells/ml, the virus titer increases in a large logarithm correspondingly, up to 1×10 9 FFU/ml, which greatly reduces the production cost and is suitable for mass production of rabies vaccines.
Description
Technical field
The present invention relates to biological product technical field, be applied to the production of rabies vaccine.
Background technology
Rabies are by the people beast that rabies virus the causes disease that takes a disease altogether, and people and all homoiothermic animal are to rabies virus susceptible all.100% death in case fall ill, unique effective method are the effective Antirabic Vaccines of inoculation.The Antirabic Vaccine who uses has HDCV (HDCV) at present; Former generation hamster kidney cell Antirabic Vaccine (PHKCV); Refining chick-embryo cell Antirabic Vaccine (PCECV); Vero cell Antirabic Vaccine (PVRV); Duck embryo Antirabic Vaccine (PDEV) etc.The Antirabic Vaccine that China produces has two kinds of PHKCV and PVRV.
Inactivated vaccine that China's approval is at present produced or attenuated live vaccine all are to prepare with mammalian cell propagative viruses method except Hepatitis B virus vaccine, mammalian cell all is important cellular matrix for viral vaccine and genetic engineering recombinant product, just begin to develop Vero cell rabies and poliomyelitis vaccine Vero cell Vaccinum Encephalitidis Epidemicae in the world as far back as the eighties, hemorrhagic fever with renal syndrome vaccine and rabies vaccine are succeeded in developing at home, the research of Vero cell SARS vaccine and Vero cell poliomyelitis vaccine has obtained bigger progress, the development technology of these developing vaccines empirical evidence China mammalian cell vaccines reaches its maturity, just cultivation scale and equipment are relatively backward, still rest on the level that rolling bottle or cell factory are cultivated, western developed countries such as America and Europe mainly are to adopt bioreactor High Density Cultivation animal cell to produce virus vaccine, it is good to have quality, the output height, low cost and other advantages, be easy to monitor good reproducibility in process of production, meet the GMP management expectancy." Chinese biological technical industry development report " book is pointed out: the cell large scale culture technique has become China's biotech drug industrialization " threshold " of difficult leap as technology the most key in the bio-pharmaceuticals.
The kind of the clear common animals cell biological reactor of order:
Gas-lifting type (airlift) bioreactor;
Hollow fiber conduit (hollow-fiber) bioreactor;
Fluid bed (fluidiz-bed) bioreactor (FBR);
Agitator tank (stir-tank) bioreactor (STR);
Fixed bed (fixed-bed) bioreactor;
Packed bed (packed-bed) bioreactor.
Microcarrier: be meant diameter at 60-250 μ m, can be applicable to the microballon of attached cell growth.Generally form by natural glucosan or various synthetic polymer.Since the first kind microcarrier appearance of Van Wezel with DEAE-Sephadex A 50 developments, the type of the microcarrier commodity of selling on the international market has reached more than tens kinds, comprises liquid microcarrier, macropore gelatin microcarrier, polystyrene microcarrier, PHEMA microcarrier, chitin microcarrier, polyurethane foam microcarrier, alginate jelly microcarrier and magnetic microcarrier etc.The convenience goods microcarrier has three kinds: Cytodex1,2,3, Cytopore and Cytoline.It is the most rising a kind of animal cell large-scale culture technique of generally acknowledging at present that microcarrier is cultivated, and it has the advantage of suspension culture and adhere-wall culture concurrently, amplifies easily.Microcarrier is cultivated to be widely used in and is cultivated all kinds cell at present, produces vaccine, protein product, as 293 cells, sarcoplast, Vero cell, Chinese hamster ovary celI.
The Chinese patent publication number is the patent application of CN102000326, a kind of Antirabic Vaccine's of production method is disclosed, the used cultured cell of this patent is a human diploid cell, human diploid cell is owing to be subjected to generation restriction and cell poor growth, so it is longer to be used for the production cycle of rabies vaccine, viral harvest yield is lower.The Chinese patent publication number is the patent application of CN101716341A, discloses a kind of human diploid cell inactivated rabies vaccine and preparation method thereof, and the used cell of this patent also is a human diploid cell, and the rabies virus strain is the PM strain.The Chinese patent publication number is applying for a patent of CN1390604, the method of large-scale continuous production of virus vaccine is disclosed, the used bioreactor of this patent is CelligenPlus, scale only is 5.0 ~ 7.5L, be not suitable for large-scale rabies vaccine production, and carrier is polyester slice Fibra-Cel Disks.
Summary of the invention
The invention provides a kind of applying biological reactor and chip carrier and produce the method for rabies vaccine, with solve the production cycle that exists in the present rabies vaccine production long, viral harvest yield is lower, is not suitable for the problem of large-scale production.
The technical scheme that the present invention takes is: comprise the following steps:
⑴ be carrier with BioNOC II cell culture disks in the Celligen 310 type 14L bioreactors with the basket stirring system of fixed bed, Cultivation of Vero, and inoculum density is 0.5～2 * 10
6Cells/ml;
⑵ grow into 5 * 10 at cell
7Individual/as during ml,, to make virus infected cell in 1:100～1:500 ratio inoculation aGV or CTN-1V rabies virus;
⑶ conditioned reaction device control parameter: CO
2Pressure is 5%～10%, pH to 7.4～7.8, temperature be 30 ℃～35 ℃, dissolved oxygen 20%～90%, concentration of glucose 25～40uM, stir speed (S.S.) at 20～100rpm, gather in the crops viral liquid;
⑷ carry out steps such as the concentrating of virus, deactivation, purification, lyophilizing and make rabies vaccine.
Owing to the raising of national standard, the production of rabies vaccine all is tending towards the applying biological reactor produces, so that every index reaches the Chinese Pharmacopoeia standard at present.Bioreactor and kind of carrier are various in the market, wherein the bioreactor quality of producing with U.S. NBS company is good, technical merit is higher, use more extensive, the corresponding supporting polyester slice Fibra-Cel Disks of U.S. NBS company that uses is also more, but the present invention finds to use the BioNOC II cell culture disks Cultivation of Vero effect of CELLIGEN 310 type 14L bioreactors collocation Taiwan Race space Fine born of the same parents Science and Technology Co., Ltd. of NBS company in the great deal of experimental process best, and cell density can reach 5 * 10
8/ ml, virus titer can reach 1 * 10
9FFU/ml, this cell density and virus titer are to use other carriers to be beyond one's reach under equal condition of culture, follow-up purification work has directly been simplified in this discovery, has solved two technological difficulties during rabies vaccine is produced: vaccine potency and host cell DNA residual quantity.
1, the present invention uses the Vero cell as the Virus culture cell.
The Vero cell is the wide spectrum host cell, at home by state approval as the production of vaccine cell, be widely used in the production of multiple vaccine, and the speed of growth is fast than human diploid cell, be more suitable in being applied to large-scale production of vaccine;
2, the used rabies virus strain of the present invention is aGV strain or CTN-1V strain.
Producing with vaccine strain is the key of producing the high-quality vaccine, the vaccine strain that China uses is mainly aG strain (rabies vaccine of hamster kidney cell), the CTN-1V strain, two vaccine strains all belong to the gene I type, the aGV strain that the present invention adapts to is 92% with PV strain homology, with CVS relatively, homology is 91%, and with street strain relatively is 84%.This seed culture of viruses adopts mice encephalocoele attack toxicity test method to record virulence can reach 8.2lgLD
50/ ml, the rabies virus that can guarantee to prepare higher titre like this is used for the production of vaccine, for subsequent purification work lays the foundation.
3, the present invention uses the CELLIGEN 310 type 14L biological reactor for cell culture of U.S. NBS company.
CELLIGEN 310 type 14L biological reactor for cell culture are state-of-the-art in the world at present animal and plant cell culture systems, are characterized in: be fit to adhere-wall culture cell and suspension cell; Be fit to various zooblasts, insect cell and plant cell etc.; The incubation of perfusion type; Low (nothing) shearing force stirs, no bubble ventilation; Detect cell concentration in real time by continuing to detect oxygen uptake rate; Its perfusion type is cultivated and is belonged to continuous culture, and cell is retained in the reactor assembly, constantly adds fresh culture in the time of the results culture fluid.The major advantage that perfusion type is cultivated is that the culture medium of continous pouring can provide the sufficient nutrition composition, and takes away metabolite.Simultaneously, cell is retained in the reactor, can reach very high cell density.Compare with additive method, the virus yield that perfusion type is cultivated can improve an order of magnitude, and can reduce labour force's consumption greatly.
The prior art scale only is 5.0 ~ 7.5L, and single virus results liquid measure is less, and the present invention is 14L, is more suitable for producing.
4, the present invention uses the BioNOC II cell culture disks of Taiwan Race space Fine born of the same parents Science and Technology Co., Ltd..
BioNOC II cell culture disks is applicable to the growth of zooblast and plant cell, its surface is made by 100% dacron element, it is the bar shaped chip carrier that 5 mm are wide and 10 mm are long, the three-dimensional porous crack of possess hydrophilic property growing surface, be fit to the cell growth, a gram BioNOC II cell culture disks can provide about 1.0 x 10
9The space of individual cell growth, just 2,400 cm
2/ g., this three-dimensional porous crack growth table mask provide hyperplasia surface area, strengthen characteristic such as cell attaching ability, can promote cell to attach, strengthen hyperplasia and diffusion, cell yield increase thereupon.The Vero cell density can reach 5 * 10 in an application of the invention
8Thereby individual/ml can prepare the rabies virus of high titre.
And use more polyester slice Fibra-Cel Disks at present, a gram only can provide 3 x 10
5The growing space of individual cell, can provide cell growth surface long-pending is 1,200cm
2/ g.
Advantage of the present invention is that cultured cells density height, virus titer and output height and production cost are low, this reactor increases and can make cell culture density reach 5 * 10 by creating good condition of culture again for the surface area of cell attachment growth by adding BioNOC II cell culture disks
8Individual/ml, virus titer becomes logarithm to increase accordingly significantly, reaches as high as 1 * 10
9FFU/ml has reduced production cost significantly, is applicable to the production in enormous quantities of rabies vaccine.
Description of drawings
Fig. 1 is for using BioNOC II cell culture disks and NBS Fibra-Cel Disks cultured cell growth curve relatively.
Fig. 2 cultivates virus titer relatively for using BioNOC II cell culture disks and NBS Fibra-Cel Disks.
The specific embodiment
Cell: African green monkey kidney (Vero) cell;
Seed culture of viruses: the aGV(virulence is greater than 7.5lgLD
50/ ml) or the CTN-1V strain;
Recovery work seed lot Vero cell goes down to posterity, increases with the 3L rolling bottle;
Celligen 310 type 14L bioreactors with the basket stirring system of fixed bed;
With BioNOC II cell culture disks is carrier.
Embodiment 1
(1) the Celligen 310 type 14L bioreactors sterilization back that will have the basket stirring system of fixed bed is assembled complete, add cell culture fluid, add BioNOC II cell culture disks microcarrier 15g/L, calf serum content 5%, adjusting PH to 7.0,33 ℃ of temperature, mixing speed speed 20rpm spend the night in the culture fluid; Inoculation is through the Vero cell of 3L rolling bottle amplification, and inoculum density is 0.5 * 10
6Cells/ml regulates CO
2Pressure is 5%, pH to 7.2, and temperature is that 35 ℃, dissolved oxygen 20%, concentration of glucose 25uM, stir speed (S.S.) are at 20rpm;
(2) increase to 5 * 10 when cell density
7Cells/ml inoculation aGV virus, inoculative proportion is 1:100,
(3) detect parameters such as glucose and lactic acid content by biochemistry analyzer, conditioned reaction device control parameter: CO
2Pressure is 5%, pH to 7.4, and temperature is that 30 ℃, dissolved oxygen 20%, concentration of glucose 25uM, stir speed (S.S.) are at 20rpm; 24 hours sampling and measuring virus virulences in every interval and antigenic content carry out the virus results with this, are higher than 1 * 10 when recording virulence
7Begin to gather in the crops single virus liquid during FFU/ml, continue 15 days harvest time, and wherein 5 days virus virulences continue 1 * 10
8FFU/ml, virulence drops to 1 * 10
6Stop results during FFU/ml;
(4) the viral liquid of results merges, ultrafiltration and concentration, deactivation, purification, lyophilizing, is distributed into the rabies vaccine finished product.
(1) the Celligen 310 type 14L bioreactors sterilization back that will have the basket stirring system of fixed bed is assembled complete, add cell culture fluid, add BioNOC II cell culture disks microcarrier 18g/L, calf serum content 8%, adjusting PH to 7.2,35 ℃ of temperature, mixing speed speed 35rpm spend the night in the culture fluid; Inoculation is through the Vero cell of 3L rolling bottle amplification, and inoculum density is 1.3 * 10
6Cells/ml regulates CO
2Pressure is 8%, pH to 7.3, and temperature is that 36 ℃, dissolved oxygen 55%, concentration of glucose 35uM, stir speed (S.S.) are at 60rpm;
(2) increase to 5 * 10 when cell density
7Cells/ml inoculation aGV virus, inoculative proportion is 1:300,
(3) detect parameters such as glucose and lactic acid content by biochemistry analyzer, conditioned reaction device control parameter: CO
2Pressure is 8%, pH to 7.6, and temperature is that 33 ℃, dissolved oxygen 55%, concentration of glucose 35uM, stir speed (S.S.) are at 60rpm; 24 hours sampling and measuring virus virulences in every interval and antigenic content carry out the virus results with this, are higher than 1 * 10 when recording virulence
7Begin to gather in the crops single virus liquid during FFU/ml, continue 15～20 days harvest time, and wherein 5～10 days virus virulences continue 1 * 10
8～1 * 10
9FFU/ml, virulence drops to 1 * 10
6Stop results during FFU/ml;
(4) the viral liquid of results merges, ultrafiltration and concentration, deactivation, purification, lyophilizing, is distributed into the rabies vaccine finished product.
Embodiment 3
(1) the Celligen 310 type 14L bioreactors sterilization back that will have the basket stirring system of fixed bed is assembled complete, add cell culture fluid, add BioNOC II cell culture disks microcarrier 20g/L, calf serum content 10%, adjusting PH to 7.4,37 ℃ of temperature, mixing speed speed 50rpm spend the night in the culture fluid; Inoculation is through the Vero cell of 3L rolling bottle amplification, and inoculum density is 2 * 10
6Cells/ml regulates CO
2Pressure is 10%, pH to 7.4, and temperature is that 37 ℃, dissolved oxygen 90%, concentration of glucose 40uM, stir speed (S.S.) are at 100rpm;
(2) increase to 5 * 10 when cell density
7Cells/ml inoculation aGV virus, inoculative proportion is 1:500,
(3) detect parameters such as glucose and lactic acid content by biochemistry analyzer, conditioned reaction device control parameter: CO
2Pressure is 10%, pH to 7.8, and temperature is that 35 ℃, dissolved oxygen 90%, concentration of glucose 40uM, stir speed (S.S.) are at 100rpm; 24 hours sampling and measuring virus virulences in every interval and antigenic content carry out the virus results with this, are higher than 1 * 10 when recording virulence
7Begin to gather in the crops single virus liquid during FFU/ml, continue 15～20 days harvest time, and wherein 5～10 days virus virulences continue 1 * 10
8～1 * 10
9FFU/ml, virulence drops to 1 * 10
6Stop results during FFU/ml;
(4) the viral liquid of results merges, ultrafiltration and concentration, deactivation, purification, lyophilizing, is distributed into the rabies vaccine finished product.
With inoculation CTN-1V Strain in the step (2) among the embodiment 1 ~ 3.
Contrast experiment's example:
(1) prepares two Celligen 310 type 14L bioreactors, the assembling of sterilization back is complete, add cell culture fluid, add BioNOC II cell culture disks and NBS Fibra-Cel Disks microcarrier 20g/L respectively, two bioreactor culture conditions are identical, are calf serum content 10% in the culture fluid, regulate pH to 7.4,37 ℃ of temperature, mixing speed speed 50rpm spend the night; Inoculation is through the Vero cell of 3L rolling bottle amplification, and inoculum density is 2 * 10
6Cells/ml regulates CO
2Pressure is 10%, pH to 7.4, and temperature is that 37 ℃, dissolved oxygen 90%, concentration of glucose 40uM, stir speed (S.S.) are at 100rpm; The 24 hours sampling and measuring cell densities in every interval.
(2) increase to 5 * 10 when two reactor cell densities
7Inoculation aGV virus during cells/ml, inoculative proportion is 1:200.
(3) regulate two reactors control parameter, make reaction condition unanimity: CO
2Pressure is 10%, pH to 7.8, and temperature is that 35 ℃, dissolved oxygen 90%, concentration of glucose 40uM, stir speed (S.S.) are at 100rpm.The 24 hours sampling and measuring virus virulences in every interval;
Contrast and experiment:
Fig. 1 is for using BioNOC II cell culture disks and NBS Fibra-Cel Disks cultured cell growth curve relatively.
Abscissa shown in the figure is the cell culture natural law, and vertical coordinate is a cell density.The initiator cell density of using above-mentioned two kinds of carriers cultivation is 2.0 * 10
6/ ml uses BioNOC II cell culture disks cultured cells and just reached 1.0 * 10 since the 8th day
8/ ml, cell density continue to raise, and cell density continued 4.0 * 10 in the 10th day to the 20th day
8~ 5.0 * 10
8Between/the ml, and reach as high as 5.0 * 10
8/ ml, the 19th day begins that downward trend is arranged; Reached 1.0 * 10 and use NBS Fibra-Cel Disks cultured cells since the 9th day
8/ ml, but cell density increasess slowly, and cell density continued 2.0 * 10 in the 11st day to the 20th day
8~ 4.0 * 10
8Between/the ml, and the highest reach 4.0 * 10
8/ ml.
Fig. 2 cultivates virus titer relatively for using BioNOC II cell culture disks and NBS Fibra-Cel Disks.
Abscissa shown in the figure is the Virus culture natural law, and vertical coordinate is a virus titer, and virus titer adopts immunofluorescence to detect.The initial virus titer of using above-mentioned two kinds of carriers cultivation is 6.0 * 10
6FFU/ml, the virus titer of using BioNOC II cell culture disks cultivation just reached 2.0 * 10 since the 10th day
8FFU/ml, and the lasting rising of virus titer reached the highest by 1.1 * 10 by the 13rd day
9FFU/ml descends afterwards successively, reduces to 10 to the 17th day virus titer
7FFU/ml; And the virus titer of using NBS Fibra-Cel Disks cultivation reached 2.0 * 10 since the 12nd day
8FFU/ml reached the highest by 6.0 * 10 by the 14th day
8FFU/ml descends afterwards successively, reduces to 10 to the 17th day virus titer
7FFU/ml.
Contrast experiment's conclusion:
More than the explanation of two groups of contrast tests to use BioNOC II cell culture disks all high more a lot of than using NBS Fibra-Cel Disks cultured cells density and virus virulence, and the persistent period of cell high density and viral high titre is all longer.Be more suitable for carrying out the production of rabies vaccine than using NBS Fibra-Cel Disks so use BioNOC II cell culture disks in the Celligen of NBS 310 type 14L bioreactors.
Claims (4)
1. applying biological reactor and chip carrier are produced the method for rabies vaccine, it is characterized in that comprising the following steps:
⑴ be carrier with BioNOC II cell culture disks in the Celligen 310 type 14L bioreactors with the basket stirring system of fixed bed, Cultivation of Vero, and inoculum density is 0.5～2 * 10
6Cells/ml;
⑵ grow into 5 * 10 at cell
7Individual/as during ml,, to make virus infected cell in 1:100～1:500 ratio inoculation aGV or CTN-1V rabies virus;
⑶ conditioned reaction device control parameter: CO
2Pressure is 5%～10%, pH to 7.4～7.8, temperature be 30 ℃～35 ℃, dissolved oxygen 20%～90%, concentration of glucose 25～40uM, stir speed (S.S.) at 20～100rpm, gather in the crops viral liquid;
⑷ carry out steps such as the concentrating of virus, deactivation, purification, lyophilizing and make rabies vaccine.
2. applying biological reactor according to claim 1 and chip carrier are produced the method for rabies vaccine, it is characterized in that:
Adding BioNOC II cell culture disks microcarrier among the step ⑴ is 15～20g/L.
3. applying biological reactor according to claim 1 and chip carrier are produced the method for rabies vaccine, it is characterized in that:
Step ⑴ regulates bioreactor parameter: CO
2Pressure is 5%～10%, pH to 7.2～7.4, and temperature is that 35 ℃～37 ℃, dissolved oxygen 20%～90%, concentration of glucose 25～40uM, stir speed (S.S.) are at 20～100rpm.
4. applying biological reactor according to claim 1 and chip carrier are produced the method for rabies vaccine, it is characterized in that:
Step ⑶ gathers in the crops viral liquid method: be higher than 1 * 10 when recording virulence
7Begin to gather in the crops single virus liquid during FFU/ml, continue 15～20 days harvest time, and wherein 5～10 days virus virulences continue 1 * 10
8～1 * 10
9FFU/ml, virulence drops to 1 * 10
6Stop results during FFU/ml.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110137119XA CN102240400A (en) | 2011-05-25 | 2011-05-25 | Method for producing rabies vaccine by applying bioreactor and sheet carrier |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110137119XA CN102240400A (en) | 2011-05-25 | 2011-05-25 | Method for producing rabies vaccine by applying bioreactor and sheet carrier |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102240400A true CN102240400A (en) | 2011-11-16 |
Family
ID=44958758
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201110137119XA Pending CN102240400A (en) | 2011-05-25 | 2011-05-25 | Method for producing rabies vaccine by applying bioreactor and sheet carrier |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102240400A (en) |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102552899A (en) * | 2012-03-07 | 2012-07-11 | 长春百克生物科技股份公司 | Method for preparing rabies vaccine by perfusion culture of bioreactor |
| CN103083654A (en) * | 2012-08-27 | 2013-05-08 | 万里明 | Process for preparing human diploid cell rabies vaccine through Celligen310 bioreactor |
| CN105296438A (en) * | 2015-10-20 | 2016-02-03 | 长春生物制品研究所有限责任公司 | Production method of virus for preparing forest encephalitis inactivated vaccine |
| CN107603956A (en) * | 2017-08-24 | 2018-01-19 | 乾元浩生物股份有限公司 | A kind of aviadenovirus scraps of paper carrier tidal type suspension culture method |
| CN107937355A (en) * | 2017-11-23 | 2018-04-20 | 长春西诺生物科技有限公司 | The method that NBS320 type bioreactor fixed bed perfusion cultures prepare dG plants of hydrophobin liquid of RABV |
| CN112342185A (en) * | 2020-11-11 | 2021-02-09 | 深圳市卫光生物制品股份有限公司 | Bioreactor process for chick embryo cell rabies virus |
| CN115044529A (en) * | 2022-05-11 | 2022-09-13 | 中国医学科学院医学生物学研究所 | Method for producing virus vaccine by large-scale culture of human diploid cells |
| CN116656628A (en) * | 2023-07-31 | 2023-08-29 | 深圳市卫光生物制品股份有限公司 | Preparation method of poxvirus vector vaccine |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1966076A (en) * | 2006-11-14 | 2007-05-23 | 广州市嘉合生物技术有限公司 | Mass-production method of hydrophobic vaccine |
| CN101851610A (en) * | 2010-03-30 | 2010-10-06 | 洛阳普莱柯生物工程有限公司 | Method for producing rabies virus antigens for animals at a large scale |
-
2011
- 2011-05-25 CN CN201110137119XA patent/CN102240400A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1966076A (en) * | 2006-11-14 | 2007-05-23 | 广州市嘉合生物技术有限公司 | Mass-production method of hydrophobic vaccine |
| CN101851610A (en) * | 2010-03-30 | 2010-10-06 | 洛阳普莱柯生物工程有限公司 | Method for producing rabies virus antigens for animals at a large scale |
Non-Patent Citations (1)
| Title |
|---|
| JEAN-CHRISTOPHE DRUGMAND，ET AL.: "《CHEMISTRY AND MATERIALS SCIENCE》", 31 December 2007, article "Growth of Mammalian and Lepidopteran Cells on BioNOC® II Disks, a Novel Macroporous Microcarrier", pages: 781-784 * |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102552899A (en) * | 2012-03-07 | 2012-07-11 | 长春百克生物科技股份公司 | Method for preparing rabies vaccine by perfusion culture of bioreactor |
| CN103083654A (en) * | 2012-08-27 | 2013-05-08 | 万里明 | Process for preparing human diploid cell rabies vaccine through Celligen310 bioreactor |
| CN105296438A (en) * | 2015-10-20 | 2016-02-03 | 长春生物制品研究所有限责任公司 | Production method of virus for preparing forest encephalitis inactivated vaccine |
| CN107603956A (en) * | 2017-08-24 | 2018-01-19 | 乾元浩生物股份有限公司 | A kind of aviadenovirus scraps of paper carrier tidal type suspension culture method |
| CN107937355A (en) * | 2017-11-23 | 2018-04-20 | 长春西诺生物科技有限公司 | The method that NBS320 type bioreactor fixed bed perfusion cultures prepare dG plants of hydrophobin liquid of RABV |
| CN112342185A (en) * | 2020-11-11 | 2021-02-09 | 深圳市卫光生物制品股份有限公司 | Bioreactor process for chick embryo cell rabies virus |
| CN112342185B (en) * | 2020-11-11 | 2022-12-02 | 深圳市卫光生物制品股份有限公司 | Bioreactor process for chick embryo cell rabies virus |
| CN115044529A (en) * | 2022-05-11 | 2022-09-13 | 中国医学科学院医学生物学研究所 | Method for producing virus vaccine by large-scale culture of human diploid cells |
| CN116656628A (en) * | 2023-07-31 | 2023-08-29 | 深圳市卫光生物制品股份有限公司 | Preparation method of poxvirus vector vaccine |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102240400A (en) | Method for producing rabies vaccine by applying bioreactor and sheet carrier | |
| CN101979518B (en) | Method for preparing pseudorabies virus | |
| CN102816732B (en) | MDCK cell line suitable for full suspended serum-free culture and its application in culturing of influenza virus and production of influenza virus vaccines | |
| CN103898041B (en) | Hybridoma Cell Culture Method | |
| CN109943471A (en) | A kind of mescenchymal stem cell filtering perfusion culture preparation facilities | |
| CN103773741A (en) | Method for preparing influenza vaccine by using cage type ventilating and stirring bioreactor | |
| CN108220227A (en) | A kind of method for the culture newcastle disease virus that suspended by the continuous cell line that suspends entirely | |
| CN114317405A (en) | Serum-free full-suspension culture type F81 cell line and construction method and application thereof | |
| CN103087995B (en) | Method of preparing foot-and-mouth disease vaccine by using BHK-21 adherent cells | |
| CN102807964A (en) | Method for scale-up culture of animal cells | |
| CN102861329A (en) | Production method of canine parvovirus inactivated vaccine by utilizing bioreactor | |
| CN105749270A (en) | Rotavirus vaccine and preparation method thereof | |
| Tyo et al. | Dense cultures of animal cells at the industrial scale | |
| CN102703374B (en) | Wall-attachment cell culture method | |
| CN102038946A (en) | Method for industrially producing pseudorabies vaccine by using bioreactor | |
| CN105838683A (en) | Method for proliferation of mink canine distemper virus by applying novel cell microcarrier | |
| CN102038947A (en) | Method for industrially producing animal rabies vaccine by utilizing bioreactor | |
| CN102038945B (en) | Method for industrially producing porcine parvovirus vaccine by using bioreactor | |
| CN102002481B (en) | Production method of porcine reproductive and respiratory syndrome virus | |
| CN102038943A (en) | Method for industrially producing porcine Japanese encephalitis (JE) vaccines by utilizing bioreactor | |
| CN102743749B (en) | Method for preparing live attenuated rubella vaccine in human diploid cells by using basket-type bioreactor | |
| CN104593335B (en) | The low cells of serum free culture system Marc 145 production pig breeding and the method for respiratory syndrome CH 1R strain virus | |
| CN107794248A (en) | A kind of method with microcarrier suspension culture CDV | |
| CN112342185A (en) | Bioreactor process for chick embryo cell rabies virus | |
| CN102886043A (en) | Binary inactivated vaccine against Japanese encephalitis virus and porcine parvovirus and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20111116 |