CN102676460A - Method for vaccinating avian influenza virus through microcarrier suspension culture cell - Google Patents
Method for vaccinating avian influenza virus through microcarrier suspension culture cell Download PDFInfo
- Publication number
- CN102676460A CN102676460A CN2012101536233A CN201210153623A CN102676460A CN 102676460 A CN102676460 A CN 102676460A CN 2012101536233 A CN2012101536233 A CN 2012101536233A CN 201210153623 A CN201210153623 A CN 201210153623A CN 102676460 A CN102676460 A CN 102676460A
- Authority
- CN
- China
- Prior art keywords
- cell
- microcarrier
- inoculation
- container
- bird flu
- 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.)
- Granted
Links
Images
Landscapes
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention provides a method for vaccinating avian influenza virus through a microcarrier suspension culture cell, which comprises the following concrete steps of: selecting a master cell needed by delivering a carrier; reproducing the cell to passage in a spinner bottle; acquiring cells in the spinner bottle; vaccinating the cells onto a microcarrier; culturing the vaccinated cells in a bioreactor; collecting the microcarrier covered with the cells; vaccinating an avian influenza virus-pancreatin; culturing the avian influenza virus-pancreatin; and collecting avian influenza virus liquid. The method solves the problems that the carrier is lost, the serum cannot be cleaned clearly, the vaccination is ununiformed, or even the avian influenza virus cannot be vaccinated, and the like occur in a process that cells cultured on the microcarrier reproduce the avian influenza virus can be solved.
Description
Technical field
The present invention relates to the avian influenza vaccine technical field, particularly, relate to a kind of method of microcarrier suspension culture cell inoculation bird flu virus.
Background technology
Past, China produced influenza vaccines through the chicken embryo culture always.Need consume a large amount of chicken embryos because the chicken embryo is produced influenza vaccines, the production cycle is long, is prone to pollute; Be not easy to control; And every fertilized eggs once can only inject a virus strains, and therefore this working method efficient is very low, is unfavorable for tackling large-scale flu outbreak.Popularity is different in different areas for China's bird flu, and the diffusion of bird flu between different areas is fast; Produce avian influenza vaccine with the chicken embryo, produce how many vaccines and must make accurate plan in advance, but in general bird flu outburst, people are difficult to purchase at short notice millions of required eggs of production vaccine.For this reason, the World Health Organization, United States Government etc. all the encourage growth cell culture technology substitute present chick embryo technique runoff yield in next life influenza vaccine.
Be that matrix prepares vaccine and has no exogenous factor and pollute, be easy to large-scale production, can keep advantages such as virus antigen is stable preferably with the mammalian cell.But, only rely on the amount of animal cell culture production virus limited, therefore the carrier culture technique is applied to the production of avian influenza vaccine, will not only keep the advantage of cell cultures but also overcome the limited shortcoming of its production output undoubtedly.At present normal use two kinds of carriers are arranged, a kind of is chip carrier, a kind of in addition is spheroid carrier.Owing to need to use a small amount of pancreatin in most bird flu virus invasion cell processes.And contain the pancreatin inhibition in the serum, therefore in inoculation and breeding bird flu virus process, can not add serum.And need add serum in the carrier culturing cell process.If use the chip carrier culturing cell, its chip carrier is to be fixed in the bio-reactor, and when inoculation and breeding bird flu virus, the serum of removing in the substratum just is very easy to, and the substratum that only needs to use PBS or do not contain serum has cleaned just.And when using the microcarrier culturing cell; Situation below inoculation and breeding bird flu virus just occur: 1) owing to spherical microcarrier little (about 100 microns); Light weight is being changed substratum or is being used the phenomenon that microcarrier is lost along with solution in the process that PBS cleans more serious; 2) cell is after covering with on the microcarrier, because the interaction between cell and the cell, a plurality of microcarriers are easy to link together; 3) in the virus inoculation process, because in order to make virus inoculation liquid and carrier uniform mixing, but the speed of bio-reactor stirring again can not be too fast, otherwise influence the poisoning intrusion cell.So often cause the part microcarrier to be deposited in bio-reactor bottom easily, and cause that to connect poison inhomogeneous even do not connect viral phenomenon at all.But microcarrier culturing cell breeding bird flu virus has following advantage: 1) enlarge easily; 2) easy collecting cell.
Summary of the invention
In order to solve a series of problems that occur in the microcarrier culturing cell breeding bird flu virus process, the invention provides a kind of method of microcarrier suspension culture cell inoculation bird flu virus.This method solves that the carrier that microcarrier culturing cell breeding bird flu virus process occurs is lost, serum clean clean, to connect poison inhomogeneous even do not connect problem such as bird flu virus.
Technical scheme of the present invention is following: the method for microcarrier suspension culture cell inoculation bird flu virus comprises following concrete steps
1: select to import needed kind of cell MDCK of carrier or Vero;
2: the method that the rolling bottle propagated cell goes down to posterity: at first 1 selected kind of cell of recovery step about growth 48h, goes down to posterity according to the ratio of 1:4 in the cell bottle of T225; Passing to 1 15L rolling bottle to the cell in 4 T225 cell bottles that cover with cell then cultivates; Carrying out passage according to the ratio of 1:4 or 1:3 about 48 hours cultivates;
3: the cell in the results rolling bottle: step 2 gained cell with PBS washing 2-3 time, is added the digestion of pancreas enzyme-EDTA solution then; The 15L rolling bottle adds 100-150ml pancreas enzyme-EDTA solution, and the rolling bottle of other type adds the digestion of pancreas enzyme-EDTA solution, collecting cell then according to similar ratio;
4: after the cell counting of results, add the ratio of 5-20 cell according to each microcarrier and carry out cell inoculation; Add 2-10 gram microcarrier in every liter of substratum;
5: the bioreactor culture inoculating cell:
The amount of bio-reactor placement carrier is the 2-10 grams per liter; The substratum of bioreactor culture cell is MEM (Hyclone); The serum working concentration is 7-10%; CO
2Air flow is 5-10%; Dissolved oxygen amount is the saturated dissolved oxygen amount of the solution of 40-60%; Temperature is 37 ℃; Stirring velocity is 30-55 rev/min, and pH is 6.8-7.4; The above microcarrier of 90-95% can reach the requirement of inoculation bird flu virus during 48-72h behind the cell inoculation;
6: after step 5 cultured cells reaches the requirement of inoculation bird flu virus; Carry out following step: under the state that stirs,, transfer to the related microcarrier of the substratum in the bio-reactor together in the container that is placed with 150-250 order stainless steel mesh through the principle of siphon; This container top contains three through holes; Can temperature control 37 ℃, the stainless steel mesh that closely is fixed together with container is equipped with in the container bottom, and a discharge opeing outlet is contained in the bottom of this container; Substratum in the container is drained through the discharge opeing outlet; Be collected in the microcarrier in the container with the PBS washing then, and discharge PBS through the bottom discharge opeing outlet of container;
7: close the discharge opeing outlet then; Joining the collection of step 6 gained to bird flu virus-pancreatin inoculation liquid has in the container of microcarrier; The bottom valve of while connection container and a hole of top Y-tube; And use peristaltic pump slowly pump bird flu virus-pancreatin inoculation liquid make it cover with on the microcarrier of cell circulating slowly so that evenly inoculation of virus, thereby avoided that direct inoculation virus causes the virus inoculation heterogeneity in bio-reactor; In addition, when in bio-reactor, inoculating, if stirring velocity is slow excessively, many microcarriers are deposited in the bio-reactor bottom, cause the virus inoculation heterogeneity;
8: after the cell inoculation virus; The container that microcarrier is housed to step 7 gained adds the MEM substratum that does not contain serum; And utilize the principle of siphon microcarrier to be joined in the bio-reactor together with substratum through siphon; Carry out viral proliferation then, the condition that virus is bred in bio-reactor is: substratum is MEM (Hyclone); CO
2Air flow is 5-10%; Dissolved oxygen amount is the saturated dissolved oxygen amount of the solution of 40-60%; Temperature is 37 ℃.Stirring velocity is 30-45 rev/min, and pH is 6.5-7.2;
9: collect the avian influenza venom.
Preferably, in the step 6, collect the microcarrier covered with cell through 150-250 order stainless steel mesh and avoid in the process of using PBS washing microcarrier, the losing of carrier.
Preferably, in the step 6, the microcarrier that the PBS washing is collected in the container repeats 2-3 time;
Preferably, in the step 7, in the viral proliferation process, when the viral proliferation medium pH be lower than 6.5 or bird flu virus HA valency be higher than 9.0, with regard to removable parts solution.
Said microcarrier is Cytodex I or hyclone.
Beneficial effect of the present invention is: the method for microcarrier suspension culture cell inoculation bird flu virus according to the invention; Can be good on the microcarrier cultured cells, inoculating bird flu virus; And obtain higher avian influenza venom of tiring; Its HA is greater than 9.0, and the microcarrier loss amount is very little, is a method that is worthy of popularization.
Description of drawings
Fig. 1, the cell inoculation upgrowth situation behind 12h on the microcarrier;
Fig. 2, the cell inoculation upgrowth situation behind 24h on the microcarrier;
Fig. 3, the cell inoculation upgrowth situation behind 48h on the microcarrier;
Fig. 4, the cell inoculation upgrowth situation behind 72h on the microcarrier.
Embodiment
Embodiment 1: microcarrier bio-reactor suspension culture breeding H5 subtype avian influenza inactivated vaccine
1: select to import needed kind of cell MDCK of carrier;
2: the method that the rolling bottle propagated cell goes down to posterity: at first 1 selected kind of cell MDCK of recovery step about growth 48h, goes down to posterity according to the ratio of 1:4 in the cell bottle of T225; Passing to 1 15L rolling bottle to the cell in 4 T225 cell bottles that cover with cell then cultivates; Carrying out passage according to the ratio of 1:4 about 48 hours cultivates; The substratum that uses in this process is MEM, and serum is foetal calf serum, and usage quantity is 7%;
3: the cell in the results rolling bottle: with PBS washing 3 times, adding concentration then is 0.25% pancreas enzyme-EDTA solution 125ml digestion, collecting cell with step 2 gained cell;
4: after the cell counting of results, the microcarrier of selecting for use is Cytodex I, adds the ratio of 20 cells according to each microcarrier and carries out cell inoculation; Add 4 gram microcarriers in every liter of substratum; Behind the cell inoculation at 12h, 24h, 48h, the upgrowth situation of 72h sampling observation of cell on microcarrier, shown in Fig. 1-4.
5: the condition of bioreactor culture cell is:
Select the NBS bio-reactor of 7.5L for use, its working volume is 5L, and the substratum of culturing cell is MEM (Hyclone); The serum working concentration is 7%; CO
2Air flow is 10%; Dissolved oxygen amount is 60% the saturated dissolved oxygen amount of solution; Temperature is 37 ℃; Stirring velocity is 40 rev/mins, and pH is 7.4; Incubation time is 60h;
6: after step 5 cultured cells reaches the requirement of inoculation bird flu virus; Carry out following step: under the state that stirs,, transfer to the related microcarrier of the substratum in the bio-reactor together in the container that is placed with 150 order stainless steel meshs through the principle of siphon; This container top contains three through holes; Can temperature control 37 ℃, the stainless steel mesh that closely is fixed together with container is equipped with in the container bottom, and a discharge opeing outlet is contained in the bottom of this container; Substratum in the container is drained through the discharge opeing outlet; Be collected in the microcarrier in the container with the PBS washing then, and discharge PBS through the bottom discharge opeing outlet of container;
7: close the discharge opeing outlet then; Joining the collection of step 6 gained to H5 subtype avian influenza virus-pancreatin inoculation liquid has in the container of microcarrier; The bottom valve of while connection container and a hole of top Y-tube; And use peristaltic pump slowly pump H5 subtype avian influenza virus-pancreatin inoculation liquid make it covering with on the microcarrier of cell circulating slowly, so that evenly inoculation of virus;
8: after the cell inoculation virus; The container that microcarrier is housed to step 7 gained adds the MEM substratum that does not contain serum; And utilize the principle of siphon microcarrier to be joined in the bio-reactor together with substratum through siphon; Carry out viral proliferation then, the condition that the H5 subtype avian influenza virus is bred in bio-reactor is: substratum is MEM (Hyclone); CO
2Air flow is 5-10%; Dissolved oxygen amount is 40% the saturated dissolved oxygen amount of solution; Temperature is 37 ℃.Stirring velocity is 40 rev/mins, and pH is 7.2;
9: collect the H5 subtype avian influenza virus.Its H5 subtype avian influenza virus liquid Yield is 10L, and the HA valency is greater than 9.0.
Embodiment 2: microcarrier suspension culture breeding H9 subtype avian influenza virus
1: select to import needed kind of cell MDCK of carrier;
2: the method that the rolling bottle propagated cell goes down to posterity: at first 1 selected kind of cell MDCK of recovery step about growth 48h, goes down to posterity according to the ratio of 1:4 in the cell bottle of T225; Passing to 1 15L rolling bottle to the cell in 4 T225 cell bottles that cover with cell then cultivates; Carrying out passage according to the ratio of 1:3 about 48 hours cultivates; The substratum that uses in this process is MEM, and serum is foetal calf serum, and usage quantity is 10%;
3: the cell in the results rolling bottle: with PBS washing 2 times, adding concentration then is 0.25% pancreas enzyme-EDTA solution 125ml digestion, collecting cell with step 2 gained cell;
4: after the cell counting of results, the microcarrier of selecting for use is Cytodex I, adds the ratio of 5 cells according to each microcarrier and carries out cell inoculation; Add 3 gram microcarriers in every liter of substratum; Behind the cell inoculation at 12h, 24h, 48h, the upgrowth situation of 72h sampling observation of cell on microcarrier.
5: the condition of bioreactor culture cell is:
Select the NBS bio-reactor of 7.5L for use, its working volume is 5L, and the substratum of culturing cell is MEM (Hyclone); The serum working concentration is 7-10%; CO
2Air flow is 10%; Dissolved oxygen amount is 40% the saturated dissolved oxygen amount of solution; Temperature is 37 ℃; Stirring velocity is 40 rev/mins, and pH is 7.4; Incubation time is 56h;
6: after step 5 cultured cells reaches the requirement of inoculation bird flu virus; Carry out following step: under the state that stirs,, transfer to the related microcarrier of the substratum in the bio-reactor together in the container that is placed with 150 order stainless steel meshs through the principle of siphon; This container top contains three through holes; Can temperature control 37 ℃, the stainless steel mesh that closely is fixed together with container is equipped with in the container bottom, and a discharge opeing outlet is contained in the bottom of this container; Substratum in the container is drained through the discharge opeing outlet; Be collected in the microcarrier in the container with the PBS washing then, and discharge PBS through the bottom discharge opeing outlet of container;
7: close the discharge opeing outlet then; Joining the collection of step 6 gained to H9 subtype avian influenza virus-pancreatin inoculation liquid has in the container of microcarrier; The bottom valve of while connection container and a hole of top Y-tube; And use peristaltic pump slowly pump H9 subtype avian influenza virus-pancreatin inoculation liquid make it covering with on the microcarrier of cell circulating slowly, so that evenly inoculation of virus;
8: after the cell inoculation virus; The container that microcarrier is housed to step 7 gained adds the MEM substratum that does not contain serum; And utilize the principle of siphon microcarrier to be joined in the bio-reactor together with substratum through siphon; Carry out viral proliferation then, the condition that the H9 subtype avian influenza virus is bred in bio-reactor is: substratum is MEM (Hyclone); CO
2Air flow is 10%; Dissolved oxygen amount is 40% the saturated dissolved oxygen amount of solution; Temperature is 37 ℃.Stirring velocity is 40 rev/mins, and pH is 6.5-7.2;
9: collect the H5 subtype avian influenza virus.Its avian influenza venom Yield is 9L, and the HA valency is greater than 9.0.
Above content is to combine concrete preferred implementation to the further explain that the present invention did, and can not assert that practical implementation of the present invention is confined to these explanations.For the those of ordinary skill of technical field under the present invention, under the prerequisite that does not break away from the present invention's design, its framework form can be flexible and changeable, can the subseries product.Just make some simple deduction or replace, all should be regarded as belonging to the scope of patent protection that the present invention is confirmed by claims of being submitted to.
Claims (6)
1. the method for microcarrier suspension culture cell inoculation bird flu virus is characterized in that, comprises following concrete steps
1: select to import needed kind of cell MDCK of carrier or Vero;
2: the method that the rolling bottle propagated cell goes down to posterity: at first 1 selected kind of cell of recovery step about growth 48h, goes down to posterity according to the ratio of 1:4 in the cell bottle of T225; Passing to 1 15L rolling bottle to the cell in 4 T225 cell bottles that cover with cell then cultivates; Carrying out passage according to the ratio of 1:4 or 1:3 about 48 hours cultivates;
3: the cell in the results rolling bottle: step 2 gained cell with PBS washing 2-3 time, is added the digestion of pancreas enzyme-EDTA solution then; The 15L rolling bottle adds 100-150ml pancreas enzyme-EDTA solution, and the rolling bottle of other type adds the digestion of pancreas enzyme-EDTA solution, collecting cell then according to similar ratio;
4: after the cell counting of results, add the ratio of 5-20 cell according to each microcarrier and carry out cell inoculation; Add 2-10 gram microcarrier in every liter of substratum;
5: the bioreactor culture inoculating cell:
The amount of bio-reactor placement carrier is the 2-10 grams per liter; The substratum of bioreactor culture cell is MEM (Hyclone); The serum working concentration is 7-10%; CO
2Air flow is 5-10%; Dissolved oxygen amount is the saturated dissolved oxygen amount of the solution of 40-60%; Temperature is 37 ℃; Stirring velocity is 30-55 rev/min, and pH is 6.8-7.4; The above microcarrier of 90-95% can reach the requirement of inoculation bird flu virus during 48-72h behind the cell inoculation;
6: after step 5 cultured cells reaches the requirement of inoculation bird flu virus; Carry out following step: under the state that stirs,, transfer to the related microcarrier of the substratum in the bio-reactor together in the container that is placed with 150-250 order stainless steel mesh through the principle of siphon; This container top contains three through holes; Can temperature control 37 ℃, the stainless steel mesh that closely is fixed together with container is equipped with in the container bottom, and a discharge opeing outlet is contained in the bottom of this container; Substratum in the container is drained through the discharge opeing outlet; Be collected in the microcarrier in the container with the PBS washing then, and discharge PBS through the bottom discharge opeing outlet of container;
7: close the discharge opeing outlet then; Joining the collection of step 6 gained to bird flu virus-pancreatin inoculation liquid has in the container of microcarrier; The bottom valve of while connection container and a hole of top Y-tube; And use peristaltic pump slowly pump bird flu virus-pancreatin inoculation liquid make it covering with on the microcarrier of cell circulating slowly, so that evenly inoculation of virus;
8: after the cell inoculation virus; The container that microcarrier is housed to step 7 gained adds the MEM substratum that does not contain serum; And utilize the principle of siphon microcarrier to be joined in the bio-reactor together with substratum through siphon; Carry out viral proliferation then, the condition that virus is bred in bio-reactor is: substratum is MEM (Hyclone); CO
2Air flow is 5-10%; Dissolved oxygen amount is the saturated dissolved oxygen amount of the solution of 40-60%; Temperature is 37 ℃;
Stirring velocity is 30-45 rev/min, and pH is 6.5-7.2;
9: collect the avian influenza venom.
2. the method for microcarrier suspension culture cell inoculation bird flu virus as claimed in claim 1 is characterized in that, in the step 6, collects the microcarrier that has covered with cell through 150-250 order stainless steel mesh.
3. the method for microcarrier suspension culture cell inoculation bird flu virus as claimed in claim 1 is characterized in that, in the step 6, the microcarrier that is collected in the container with the PBS washing repeats 2-3 time.
4. the method for microcarrier suspension culture cell inoculation bird flu virus as claimed in claim 1; It is characterized in that; In the step 7, in the bird flu virus seeded process, microcarrier and cell are under immobilising situation; Through peristaltic pump slowly pump bird flu virus-pancreatin inoculation liquid make it covering with on the microcarrier of cell circulating slowly, so that evenly inoculation of virus.
5. the method for microcarrier suspension culture cell inoculation bird flu virus as claimed in claim 1; It is characterized in that, in the step 7, in the viral proliferation process; When the viral proliferation medium pH be lower than 6.5 or bird flu virus HA valency be higher than 9.0, with regard to removable parts solution.
6. the method for microcarrier suspension culture cell inoculation bird flu virus as claimed in claim 1 is characterized in that, said microcarrier is Cytodex I or hyclone.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210153623.3A CN102676460B (en) | 2012-06-19 | 2012-06-19 | Method for vaccinating avian influenza virus through microcarrier suspension culture cell |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210153623.3A CN102676460B (en) | 2012-06-19 | 2012-06-19 | Method for vaccinating avian influenza virus through microcarrier suspension culture cell |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102676460A true CN102676460A (en) | 2012-09-19 |
| CN102676460B CN102676460B (en) | 2014-02-26 |
Family
ID=46809016
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210153623.3A Active CN102676460B (en) | 2012-06-19 | 2012-06-19 | Method for vaccinating avian influenza virus through microcarrier suspension culture cell |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102676460B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103160457A (en) * | 2013-04-11 | 2013-06-19 | 山东信得动物疫苗有限公司 | Large-scaled cell-culturing proliferating method and method for using proliferated cell in virus production |
| CN104073470A (en) * | 2014-06-30 | 2014-10-01 | 中国农业科学院上海兽医研究所 | Spinner-flask culture method for H9N2 subtype of avian influenza virus |
| CN107326016A (en) * | 2017-06-26 | 2017-11-07 | 中国水产科学研究院珠江水产研究所 | A kind of microcarrier suspension culture CPB cells production mandarin fish infectious spleen and kidney necrosis virus and the method for mandarin fish rhabdovirus |
| CN109797101A (en) * | 2017-11-17 | 2019-05-24 | 北京中原合聚经贸有限公司 | A kind of cell reactor microcarrier cell harvest and inoculate amplification method |
| CN115094023A (en) * | 2022-07-04 | 2022-09-23 | 无锡多宁生物科技有限公司 | MDCK cell microcarrier culture and suspension domestication process |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101818131A (en) * | 2010-02-01 | 2010-09-01 | 成都天邦生物制品有限公司 | Production method for influenza virus vaccines |
| CN101955915A (en) * | 2010-02-01 | 2011-01-26 | 成都天邦生物制品有限公司 | Method for producing influenza virus vaccine |
| CN102127524A (en) * | 2010-12-20 | 2011-07-20 | 中山大学 | Method for proliferating avian influenza viruses in bioreactor with cell carrier |
-
2012
- 2012-06-19 CN CN201210153623.3A patent/CN102676460B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101818131A (en) * | 2010-02-01 | 2010-09-01 | 成都天邦生物制品有限公司 | Production method for influenza virus vaccines |
| CN101955915A (en) * | 2010-02-01 | 2011-01-26 | 成都天邦生物制品有限公司 | Method for producing influenza virus vaccine |
| CN102127524A (en) * | 2010-12-20 | 2011-07-20 | 中山大学 | Method for proliferating avian influenza viruses in bioreactor with cell carrier |
Non-Patent Citations (1)
| Title |
|---|
| 史爱华等: "H9N2亚型禽流感病毒在MDCK细胞中增殖最佳条件研究", 《动物医学进展》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103160457A (en) * | 2013-04-11 | 2013-06-19 | 山东信得动物疫苗有限公司 | Large-scaled cell-culturing proliferating method and method for using proliferated cell in virus production |
| CN103160457B (en) * | 2013-04-11 | 2014-10-01 | 山东信得动物疫苗有限公司 | Large-scaled cell-culturing proliferating method and method for using proliferated cell in virus production |
| CN104073470A (en) * | 2014-06-30 | 2014-10-01 | 中国农业科学院上海兽医研究所 | Spinner-flask culture method for H9N2 subtype of avian influenza virus |
| CN107326016A (en) * | 2017-06-26 | 2017-11-07 | 中国水产科学研究院珠江水产研究所 | A kind of microcarrier suspension culture CPB cells production mandarin fish infectious spleen and kidney necrosis virus and the method for mandarin fish rhabdovirus |
| CN109797101A (en) * | 2017-11-17 | 2019-05-24 | 北京中原合聚经贸有限公司 | A kind of cell reactor microcarrier cell harvest and inoculate amplification method |
| CN115094023A (en) * | 2022-07-04 | 2022-09-23 | 无锡多宁生物科技有限公司 | MDCK cell microcarrier culture and suspension domestication process |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102676460B (en) | 2014-02-26 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102676460B (en) | Method for vaccinating avian influenza virus through microcarrier suspension culture cell | |
| CN101979517B (en) | Method for producing influenza viruses in large scale by using bioreactor | |
| CN101804203A (en) | Method for mass production of pseudorabies virus vaccine | |
| CN103773741B (en) | The method that cage air agitation bioreactor prepares influenza vaccines | |
| CN102100910B (en) | Method for producing viral vaccines | |
| CN102690791A (en) | Method for generating porcine pseudorabies virus by culturing ST cell in microcarrier of bioreactor | |
| CN107460156A (en) | The serum-free strain of suspension mdck cell and its application in influenza virus is produced entirely | |
| CN106085946A (en) | Can be with the Pig testicular cell strain ST S of suspension culture and preparation method thereof and application | |
| CN102091329A (en) | Preparation method of inactivated porcine parvovirus vaccine and product thereof | |
| CN104001167A (en) | Process for preparing avian influenza inactivated vaccine by full suspended culture cells and product | |
| CN104726392B (en) | A kind of method of the suspension mammalian cell system preparing free serum culture and its cell line prepared and application | |
| CN101831412A (en) | Method for producing porcine reproductive and respiratory syndrome viruses (PRRSV) in large scale | |
| CN102807964A (en) | Method for scale-up culture of animal cells | |
| CN103520715B (en) | Method for producing porcine circovirus II type inactivated vaccine by utilizing WAVE bioreactor | |
| CN102732487B (en) | Method for large scale cultivation in bioreactor | |
| CN103087995A (en) | Method of preparing foot-and-mouth disease vaccine by using BHK-21 adherent cells | |
| CN105838683A (en) | Method for proliferation of mink canine distemper virus by applying novel cell microcarrier | |
| CN102409020A (en) | Non woven/polyester fiber carriers for culturing cells and use method thereof | |
| CN102657859A (en) | Method for producing avian influenza vaccine by using cells cultured by spherical microcarrier bioreactor | |
| CN102002482B (en) | Method for producing PRRS (Porcine Reproductive and Respiratory Syndrome) viruses | |
| CN102886043B (en) | Binary inactivated vaccine against Japanese encephalitis virus and porcine parvovirus and preparation method thereof | |
| CN102002481A (en) | Production method of porcine reproductive and respiratory syndrome virus | |
| CN104436184A (en) | Cell production method for influenza virus vaccine | |
| CN103614344A (en) | Method for amplifying porcine circovirus type 2 by applying bioreactor and flaky vector | |
| CN104630159A (en) | Method for producing hog cholera C-strain virus by culturing ST Cells in low serum |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C41 | Transfer of patent application or patent right or utility model | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20160622 Address after: 526238 Guangdong Province Wang Zhaoqing City Economic Development Zone Industrial Park abundance Patentee after: Zhaoqing Dahuanong Biological Pharmaceutical Co., Ltd. Patentee after: GUANGDONG WENS DAHUANONG BIOTECHNOLOGY CO., LTD. Address before: 526000 Guangdong Province Wang Zhaoqing City Economic Development Zone Industrial Park abundance Patentee before: Zhaoqing Dahuanong Biological Pharmaceutical Co., Ltd. Patentee before: Guangdong Dahuanong Animal Health Products Co., Ltd. |