CN103536913A - Method for producing avian influenza virus bivalent inactivated vaccine by use of MDCK cell line - Google Patents
Method for producing avian influenza virus bivalent inactivated vaccine by use of MDCK cell line Download PDFInfo
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- CN103536913A CN103536913A CN201310452808.9A CN201310452808A CN103536913A CN 103536913 A CN103536913 A CN 103536913A CN 201310452808 A CN201310452808 A CN 201310452808A CN 103536913 A CN103536913 A CN 103536913A
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Abstract
The invention provides a method for producing avian influenza virus (H5 subtype and H9 subtype) bivalent inactivated vaccine by use of a MDCK cell line. The method comprises the following steps: (1) passage and culture of the MDCK cell line; (2) virus inoculation and reproduction; (3) inactivation and harvesting of virus liquid; and (4) preparation of the vaccine. The method for producing avian influenza virus inactivated vaccine by use of the MDCK cell line has the advantages that exogenous factor pollution is avoided, large-scale production is easy to realize, the virus antigen stability can be maintained well and the like; and moreover, the development period and production period of the vaccine can be shortened, and the ability of emergency supply of vaccine in outburst epidemic situation is realized.
Description
Technical field
The present invention relates to biological product technical field, specifically, relating to a kind of employing mdck cell is the method for producing bird flu virus (H5 hypotype and H9 hypotype) bivalent inactivated vaccine.
Background technology
At present, the mode of China's production bird flu virus inactivated vaccine is mainly to utilize the healthy Embryo Gallus domesticus of 9-11 age in days to cultivate.This cultural method is by virus inoculation is carried out to virus breeding in chick embryo allantoic cavity, this mode of production can not meet following China to the prevention of Bird Flu Disease and demand for control, be mainly manifested in the following aspects: (1) production cycle is long, be difficult to realize at short notice extensive production of vaccine; (2) be subject to the restriction of the raw and auxiliary materials such as Embryo Gallus domesticus; (3) production technology falls behind, labour intensive, and production cost is high.Bird flu virus (H5 hypotype+H9 hypotype) bivalent inactivated vaccine has very important effect in China's Bird Flu Disease prevention and control process; the mode of production of employing mdck cell has without exogenous factor to be polluted; be easy to large-scale production, can maintain preferably the advantages such as virus antigen is stable; and can shorten vaccine R&D cycle and production cycle; and possess the emergent ability of vaccine supply in SARS Epidemic, be the development trend that following avian influenza vaccine is produced.
Summary of the invention
The present invention is intended to overcome the defect existing in prior art, and it is the method for producing bird flu virus (H5 hypotype and H9 hypotype) bivalent inactivated vaccine that a kind of employing mdck cell is provided.
In order to realize the object of the invention, a kind of method that adopts mdck cell system to produce bird flu virus bivalent inactivated vaccine of the present invention, comprises the following steps:
(1) what mdck cell was goes down to posterity and cultivates: mdck cell system disperses through pancreas enzyme-EDTA digestion, with cell growth medium, is diluted to after finite concentration, is seeded in suitable container, goes down to posterity or virus inoculation after cell covers with;
(2) virus inoculation and breeding: after respectively H5 subtype avian influenza virus and H9 subtype avian influenza virus suitably being diluted with viral growth liquid, inoculate respectively the mdck cell that cleans (cleaning 3 times) with PBS, after the complete pathological changes of mdck cell, stop cultivating;
(3) deactivation of virus liquid and results: in above-mentioned cell culture fluid, add after formalin deactivation, gather in the crops respectively H5 subtype avian influenza virus antigen and H9 subtype avian influenza virus antigen;
(4) preparation vaccine: make water after H5 subtype avian influenza virus antigen and H9 subtype avian influenza virus antigen are sneaked into tween 80, with together emulsifying of oil phase, make finished product vaccine after packing.
Preferably, the H5 subtype avian influenza virus described in step (2) is restructuring bird flue virus H 5 N 1 subtype Re-4 strain or recombinant fowl influenza virus H5N1 hypotype Re-6 strain, and H9 subtype avian influenza virus is restructuring Avian Influenza Virus H9N2 Re-2 strain.
Preferably, the mdck cell described in step (2) is the cell of plane adhere-wall culture or adopts bioreactor to carry out the cell of suspension culture.
Preferably, the DMEM liquid that contains 90-95% volume in the cell growth medium described in step (1) and the hyclone of 5-10% volume form, pH value 7.0-7.4.
Preferably, the water described in step (4) comprises 46.5 parts of H5 subtype avian influenza virus antigens, 46.5 parts of H9 subtype avian influenza virus antigens and 7 parts of tween 80s; Oil phase comprises 93 parts of white oils, 5 parts of Arlacel-80s and 2 parts of aluminium stearate.While carrying out emulsifying, water is 1 part, 2 parts of oil phases (described umber is quality proportion).
Preferably, in step (1), cell culture condition is 37 ℃, 5%CO
2.
Preferably, the viral growth liquid described in step (2) is to contain the DMEM liquid that 5 μ g/mL TPCK process pancreatin and 0.2% bovine serum albumin component V, pH value 7.2-7.4.
Preferably, in step (2), cell culture condition is 34 ℃, 5%CO
2.
While preferably, inoculating in step (2), H5 subtype avian influenza virus inoculum concentration is MOI10
-4-10
-6, H9 subtype avian influenza virus inoculum concentration is MOI10
-5-10
-7.
Preferably, described in step (3), the concentration of formalin is 0.1%.
Preferably, in step (3), deactivation condition is 37 ℃, deactivation 24h.
The present invention at least has following advantages and beneficial effect:
The present invention adopts mdck cell system to produce bird flu virus (H5 hypotype+H9 hypotype) bivalent inactivated vaccine first, for the prevention and control of Bird Flu Disease provide new technology.Bird flu virus (H5 hypotype+H9 hypotype) bivalent inactivated vaccine occupies for a long time in the prevention and control process of the huge share ， China Bird Flu Disease in China's avian influenza vaccine market and has played important straight vital effect.Adopt mdck cell to carry out the production of bird flu virus (H5 hypotype+H9 hypotype) bivalent inactivated vaccine, improved greatly the production technology of vaccine.Be embodied in without exogenous factor and pollute, be easy to large-scale production, can maintain preferably the advantages such as virus antigen is stable, and can shorten vaccine R&D cycle and production cycle, and possess the emergent ability of vaccine supply in SARS Epidemic.
The specific embodiment
Following examples are used for illustrating the present invention, but are not used for limiting the scope of the invention.If do not specialize, the conventional means that in embodiment, technological means used is well known to those skilled in the art, the raw materials used commercial goods that is.
The percentage sign relating in the present invention " % ", if not specified, refers to mass percent; But the percentage ratio of solution, except as otherwise herein provided, refers to the grams that contains solute in 100mL solution; Percentage ratio between liquid, refers to the ratio of capacity in the time of 20 ℃.
The plane adhere-wall culture of embodiment 1 recombinant fowl influenza virus H5N1 hypotype Re-4 strain
Comprise the following steps:
(1) adopt 850cm
2spinner culture mdck cell, select the good monolayer mdck cell of cellular morphology as Virus culture cell.
(2), with 0.01mol/L, the PBS of pH7.2-7.4 cleans mdck cell 3 times.
(3) with viral growth liquid (processing the DMEM liquid of pancreatin, 0.2% bovine serum albumin component V containing the TPCK of 5 μ g/mL, pH value 7.2-7.4), recombinant fowl influenza virus H5N1 hypotype Re-4 strain is diluted to containing viral MOI10
-4-10
-6.
(4) virus liquid having diluted is seeded in the mdck cell after the middle cleaning of step (2), is placed in 34 ℃, 5%CO
2middle cultivation 2-4 days, measures viral HA and tires, and adds 0.1% formalin in 37 ℃ of deactivations 24 hours, results virus antigen.
The bioreactor suspension culture of embodiment 2 recombinant fowl influenza virus H5N1 hypotype Re-4 strains
Comprise the following steps:
(1) utilize TideCell-010 tidal type high-density cells culture systems to carry out mdck cell filling type suspension culture:
The cell (1 * 10 of TideCell-010 tidal type high-density cells culture systems inoculation 10000mL suitable concentration
6-4 * 10
6individual cell/mL).Tissue Culture Flask is connected containing the culture bag (liquid tank) of 40000mL cell growth medium and cultivated.It is the DMEM liquid of 95% percent by volume and the hyclone of 5% percent by volume that cell growth medium is cultivated, pH value 7.2-7.4.By regulating each condition of culture in cell culture system: dissolved oxygen amount is 50%-100%, CO
2concentration is 5%, and glucose content is 500-4500mg/L, pH value 7.2-7.4, and 37 ℃ of cell culture temperature, cultivate highdensity mdck cell.Be cultured to the 2nd day, to pour into the speed of 1 working volume every day, carry out filling type cultivation, cultivate altogether 4-6 days.
(2) large-scale culture of recombinant fowl influenza virus H5N1 hypotype Re-4 strain:
The cell growth medium of tidal type high-density cells culture systems is discarded, with PBS, clean after 3 times, inoculation (is 10 containing recombinant fowl influenza virus H5N1 hypotype Re-4 strain MOI containing the viral growth liquid of appropriate virus
-4-10
-6, containing the TPCK processing pancreatin of 5 μ g/mL, the DMEM liquid of 0.2% bovine serum albumin component V, pH value 7.2-7.4, Virus culture temperature is 34 ℃), continue to cultivate.
(3) 2-4 days after inoculation, measures viral HA and tires, add final concentration be 0.1% formalin after 37 ℃ of deactivation 24h, aseptic results virus antigen.
The plane adhere-wall culture of embodiment 3 recombinant fowl influenza virus H5N1 hypotype Re-6 strains
Comprise the following steps:
(1) adopt 850cm
2spinner culture mdck cell, select the good monolayer mdck cell of cellular morphology as Virus culture cell.
(2), with 0.01mol/L, the PBS of pH7.2-7.4 cleans mdck cell 3 times.
(3) with viral growth liquid (processing the DMEM liquid of pancreatin, 0.2% bovine serum albumin component V containing the TPCK of 5 μ g/mL, pH value 7.2-7.4), recombinant fowl influenza virus H5N1 hypotype Re-6 strain is diluted to containing viral MOI10
-4-10
-6.
(4) virus liquid having diluted is seeded to the mdck cell after cleaning in step (2), puts 34 ℃, 5%CO
2middle cultivation 2-4 days, measures viral HA and tires, and adds 0.1% formalin in 37 ℃ of deactivation 24h, results virus antigen.
The bioreactor suspension culture of embodiment 4 recombinant fowl influenza virus H5N1 hypotype Re-6 strains
Comprise the following steps:
(1) utilize TideCell-010 tidal type high-density cells culture systems to carry out mdck cell filling type suspension culture:
The cell (1 * 10 of TideCell-010 tidal type high-density cells culture systems inoculation 10000mL suitable concentration
6-4 * 10
6individual cell/mL).Tissue Culture Flask is connected containing the culture bag (liquid tank) of 40000mL cell growth medium and cultivated.It is the DMEM liquid of 95% percent by volume and the new-born calf serum of 5% percent by volume that cell growth medium is cultivated, pH value 7.2-7.4.By regulating each condition of culture in cell culture system: dissolved oxygen amount is 50%-100%, CO
2concentration is 5%, glucose content is 500-4500mg/L, pH value 7.2-7.4, and 37 ℃ of cell culture temperature, cultivate highdensity mdck cell.Be cultured to the 2nd day, to pour into the speed of 1 working volume every day, carry out filling type cultivation, cultivate altogether 4～6 days.
(2) large-scale culture of recombinant fowl influenza virus H5N1 hypotype Re-6 strain:
The cell growth medium of tidal type high-density cells culture systems is discarded, with PBS, clean after 3 times, inoculation (is 10 containing recombinant fowl influenza virus H5N1 hypotype Re-4 strain MOI containing the viral growth liquid of appropriate virus
-4-10
-6, containing the TPCK processing pancreatin of 5 μ g/mL, the DMEM liquid of 0.2% bovine serum albumin component V, pH value 7.2-7.4, Virus culture temperature is 34 ℃), continue to cultivate.
(3) 2-4 days after inoculation, measures viral HA and tires, add final concentration be 0.1% formalin after 37 ℃ of deactivation 24h, aseptic results virus antigen.
The plane adhere-wall culture of embodiment 5 recombinant fowl influenza virus H9N2 hypotype Re-2 strains
Comprise the following steps:
(1) adopt 850cm
2spinner culture mdck cell, select the good monolayer mdck cell of cellular morphology as Virus culture cell.
(2), with 0.01mol/L, the PBS of pH7.2-7.4 cleans mdck cell 3 times.
(3) with viral growth liquid (processing the DMEM liquid of pancreatin, 0.2% bovine serum albumin component V containing the TPCK of 5 μ g/mL, pH value 7.2-7.4), recombinant fowl influenza virus H5N1 hypotype Re-6 strain is diluted to containing viral MOI10
-5-10
-7.
(4) virus liquid having diluted is seeded to the mdck cell after cleaning in step (2), is placed in 34 ℃, 5%CO
2middle cultivation 2-4 days, measures viral HA and tires, and adds 0.1% formalin in 37 ℃ of deactivation 24h, results virus antigen.
The bioreactor suspension culture of embodiment 6 recombinant fowl influenza virus H9N2 hypotype Re-2 strains
Comprise the following steps:
(1) utilize TideCell-010 tidal type high-density cells culture systems to carry out mdck cell filling type suspension culture:
The cell (1 * 10 of TideCell-010 tidal type high-density cells culture systems inoculation 10000mL suitable concentration
6-4 * 10
6individual cell/mL).Tissue Culture Flask is connected containing the culture bag (liquid tank) of 40000mL cell growth medium and cultivated.It is the DMEM liquid of 95% percent by volume and the new-born calf serum of 5% percent by volume that cell growth medium is cultivated, pH value 7.2-7.4.By regulating each condition of culture in cell culture system: dissolved oxygen amount is 50%-100%, CO
2concentration is 5%, and glucose content is 500-4500mg/L, pH value 7.2-7.4, and 37 ℃ of cell culture temperature, cultivate highdensity mdck cell.Be cultured to the 2nd day, to pour into the speed of 1 working volume every day, carry out filling type cultivation, cultivate altogether 4～6 days.
(2) large-scale culture of recombinant fowl influenza virus H9N2 hypotype Re-2 strain:
The cell growth medium of tidal type high-density cells culture systems is discarded, with PBS, clean after 3 times, inoculation (is 10 containing recombinant fowl influenza virus H5N1 hypotype Re-4 strain MOI containing the viral growth liquid of appropriate virus
-4-10
-6, containing the TPCK processing pancreatin of 5 μ g/mL, the DMEM liquid of 0.2% bovine serum albumin component V, pH value 7.2-7.4, Virus culture temperature is 34 ℃), continue to cultivate.
(3) 2-4 days after inoculation, measures viral HA and tires, add final concentration be 0.1% formalin after 37 ℃ of deactivation 24h, aseptic results virus antigen.
In embodiment 1-6, bird flu virus cultivation results is as shown in table 1.
Table 1 bird flu virus cultivation results
The immunity test of embodiment 7 bird flu viruss (H5N1 hypotype Re-4 strain+H9N2 hypotype Re-2 strain) bivalent inactivated vaccine
Comprise the following steps:
(1) vaccine preparation: make water after 46.5 parts of recombinant fowl influenza virus H5N1 hypotype Re-4 strain antigens and 46.5 parts of recombinant fowl influenza virus H9N2 hypotype Re-2 strain antigens are sneaked into 7 parts of aseptic tween 80s; 93 parts of white oils, 5 parts of Arlacel-80s and 2 parts of aluminium stearate are fully mixed to transparent rear autoclaving and make oil phase.Get 2 parts of oil phases and 1 part of water carries out emulsifying (described umber is quality proportion), after packing, make vaccine.
(2) vaccine immunity: vaccine is injected 10 of 3-4 SPF chickens in age in week by chest muscle, and dosage of inoculation is every chicken 0.3mL.5 chickens are not injected in contrast.Inoculate after 21 days, blood sampling separation of serum, tires with bird flu virus H5 hypotype antigen and H9 hypotype Detection of antigen HI.
The immunity test of embodiment 8 bird flu viruss (H5N1 hypotype Re-6 strain+H9N2 hypotype Re-2 strain) bivalent inactivated vaccine
Comprise the following steps:
(1) vaccine preparation: make water after 46.5 parts of recombinant fowl influenza virus H5N1 hypotype Re-6 strain antigens and 46.5 parts of recombinant fowl influenza virus H9N2 hypotype Re-2 strain antigens are sneaked into 7 parts of aseptic tween 80s; 93 parts of white oils, 5 parts of Arlacel-80s and 2 parts of aluminium stearate are fully mixed to transparent rear autoclaving and make oil phase.Get 2 parts of oil phases and 1 part of water carries out emulsifying (described umber is quality proportion), after packing, make vaccine.
(2) vaccine immunity: vaccine is injected 10 of 3-4 SPF chickens in age in week by chest muscle, and dosage of inoculation is every chicken 0.3ml.5 chickens are not injected in contrast.Inoculate after 21 days, blood sampling separation of serum, tires with bird flu virus H5 hypotype antigen and H9 hypotype Detection of antigen HI.
Embodiment 7 and 8 vaccine immunity result of the test are as shown in table 2.
Table 2 vaccine immunity result of the test
Although above the present invention is described in detail with a general description of the specific embodiments, on basis of the present invention, can make some modifications or improvements it, this will be apparent to those skilled in the art.Therefore, these modifications or improvements, all belong to the scope of protection of present invention without departing from theon the basis of the spirit of the present invention.
Claims (10)
1. the method that adopts mdck cell system to produce bird flu virus bivalent inactivated vaccine, is characterized in that, comprises the following steps:
(1) what mdck cell was goes down to posterity and cultivates: mdck cell system disperses through pancreas enzyme-EDTA digestion, with cell growth medium, is diluted to after finite concentration, is seeded in suitable container, goes down to posterity or virus inoculation after cell covers with;
(2) virus inoculation and breeding: after respectively H5 subtype avian influenza virus and H9 subtype avian influenza virus suitably being diluted with viral growth liquid, inoculate respectively the mdck cell cleaning with PBS, after the complete pathological changes of mdck cell, stop cultivating;
(3) deactivation of virus liquid and results: in above-mentioned cell culture fluid, add after formalin deactivation, gather in the crops respectively H5 subtype avian influenza virus antigen and H9 subtype avian influenza virus antigen;
(4) preparation vaccine: make water after H5 subtype avian influenza virus antigen and H9 subtype avian influenza virus antigen are sneaked into tween 80, with together emulsifying of oil phase, make finished product vaccine after packing.
2. method according to claim 1, it is characterized in that, H5 subtype avian influenza virus described in step (2) is restructuring bird flue virus H 5 N 1 subtype Re-4 strain or recombinant fowl influenza virus H5N1 hypotype Re-6 strain, and H9 subtype avian influenza virus is restructuring Avian Influenza Virus H9N2 Re-2 strain.
3. method according to claim 1, is characterized in that, the mdck cell described in step (2) is the cell of plane adhere-wall culture or adopts bioreactor to carry out the cell of suspension culture.
4. method according to claim 1, is characterized in that, the DMEM liquid that contains 90-95% volume in the cell growth medium described in step (1) and the hyclone of 5-10% volume, pH value 7.0-7.4.
5. method according to claim 1, is characterized in that, the oil phase described in step (4) comprises white oil, Arlacel-80 and aluminium stearate.
6. method according to claim 1, is characterized in that, in step (1), cell culture condition is 37 ℃, 5%CO
2.
7. method according to claim 1, is characterized in that, the viral growth liquid described in step (2) is to contain the DMEM liquid that 5 μ g/mL TPCK process pancreatin and 0.2% bovine serum albumin component V, pH value 7.2-7.4.
8. method according to claim 1, is characterized in that, in step (2), cell culture condition is 34 ℃, 5%CO
2.
9. method according to claim 1, is characterized in that, while inoculating in step (2), H5 subtype avian influenza virus inoculum concentration is MOI10
-4-10
-6, H9 subtype avian influenza virus inoculum concentration is MOI10
-5-10
-7.
10. method according to claim 1, is characterized in that, described in step (3), the concentration of formalin is 0.1%, and deactivation condition is 37 ℃, deactivation 24h.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105816870A (en) * | 2016-03-24 | 2016-08-03 | 福州大北农生物技术有限公司 | Preparation method of avian influenza virus inactivating vaccine |
| CN108130315A (en) * | 2017-12-20 | 2018-06-08 | 哈药集团生物疫苗有限公司 | H3N2 hypotype swine influenza virus cell adapted strains and its inactivated vaccine and the application of preparation |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN102526718A (en) * | 2010-12-30 | 2012-07-04 | 华中农业大学 | Recombinant H5N1 (Hemagglutinin 5 Neuraminidase 1) avian influenza virus cell vaccine and application thereof |
-
2013
- 2013-09-27 CN CN201310452808.9A patent/CN103536913A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN102526718A (en) * | 2010-12-30 | 2012-07-04 | 华中农业大学 | Recombinant H5N1 (Hemagglutinin 5 Neuraminidase 1) avian influenza virus cell vaccine and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| 刘洁等: "重组禽流感病毒H5亚型二价灭活疫苗（Re-4+Re-6）实验报告", 《养殖技术顾问》 * |
| 田国彬等: "禽流感H5、H9二价灭活疫苗(H5N1 Re-1株，H9N2 Re-2株)研究及应用", 《中国畜牧兽医医学会畜牧兽医生物技术学分会暨中国免疫学会兽医免疫分会第六次研讨会论文集》 * |
| 詹丽娥等: "禽流感H5N1-H9N2亚型二价灭活疫苗的研制及应用", 《中国畜牧兽医学会家畜传染病学分会第六次全国会员代表大会暨第11次学术研讨会论文集》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105816870A (en) * | 2016-03-24 | 2016-08-03 | 福州大北农生物技术有限公司 | Preparation method of avian influenza virus inactivating vaccine |
| CN108130315A (en) * | 2017-12-20 | 2018-06-08 | 哈药集团生物疫苗有限公司 | H3N2 hypotype swine influenza virus cell adapted strains and its inactivated vaccine and the application of preparation |
| CN108130315B (en) * | 2017-12-20 | 2021-08-13 | 哈药集团生物疫苗有限公司 | H3N2 subtype swine influenza virus cell adapted strain, inactivated vaccine prepared from same and application of inactivated vaccine |
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Application publication date: 20140129 |