CN103768589A - Method for removing residual DNA in encephalitis B vaccine product by utilizing hollow fiber membrane - Google Patents
Method for removing residual DNA in encephalitis B vaccine product by utilizing hollow fiber membrane Download PDFInfo
- Publication number
- CN103768589A CN103768589A CN201210398655.XA CN201210398655A CN103768589A CN 103768589 A CN103768589 A CN 103768589A CN 201210398655 A CN201210398655 A CN 201210398655A CN 103768589 A CN103768589 A CN 103768589A
- Authority
- CN
- China
- Prior art keywords
- encephalitis
- human
- vaccine
- liquid
- fibre membrane
- 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
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention relates to a method for removing residual DNA in an encephalitis B vaccine product for human by utilizing a hollow fiber membrane. The method is characterized in that an encephalitis B vaccine obtaining liquid for human is used as a raw material, other proteins and DNA in the encephalitis B vaccine obtaining liquid for human are removed through a hollow fiber membrane microfiltration process, the obtained filtered liquid is collected and is processed by an ultrafilration concentration system to prepare an encephalitis B vaccine concentrate for human, the concentrate is inactivated, and the obtained inactivated encephalitis B vaccine liquid for human is purified by using a molecular sieve chromatography purifying system. A certain proportion of DNA residual in the product is effectively removed through the microfiltration treatment of the virus obtaining liquid by the hollow fiber membrane without influencing the biological activity of the vaccine product, so the residual quantity of DNA in the finished vaccine product reaches a qualified standard.
Description
Technical field
The present invention relates to remove in a kind of encephalitis B used for human being vaccine product the method for DNA and foreign protein, particularly relate to a kind of method of utilizing hollow-fibre membrane to remove residual DNA in encephalitis B used for human being vaccine product.The hollow-fibre membrane that the present invention mentions is hollow fiber microfiltration membrane, and its material main component is PP or PVDF etc.
Background technology
Encephalitis B used for human being vaccine is that encephalitis b virus is inoculated on applicable zooblast, after breeding, virus is discharged in culture fluid, some remaining host cells or its fragment also can be along with being mixed in culture fluid around here, through clarification, concentrated deactivation, after purification, extraction obtains in vaccine for virus of encephalitis B stock solution, can produce free host DNA and the host DNA of being combined with antigen protein, in stock solution, also contain a large amount of host proteins, concentrated and purified technique can be removed a certain proportion of free DNA, exist but still have residual DNA, particularly can remain in purification stock solution with the DNA that virus or antigen protein are combined, as not thorough in removed, this part residual DNA can be injected in human body together along with finished product vaccine, may in human body, cause untoward reaction, or have cause cancer occur possibility.
Summary of the invention
Object of the present invention is just to solve prior art above shortcomings, provides a kind of method of utilizing hollow-fibre membrane to remove residual DNA in encephalitis B used for human being vaccine product.The present invention, not affecting under the bioactive prerequisite of vaccine product, carries out microfiltration processing by hollow-fibre membrane to virus harvest liquid, effectively removes a certain proportion of DNA remaining in wherein, thereby makes finished product Vaccines DNA residual quantity reach criterion of acceptability.
The technical scheme that the present invention provides is: this method of utilizing hollow-fibre membrane to remove residual DNA in encephalitis B used for human being vaccine product, be characterized in: take encephalitis B used for human being vaccine harvest liquid as raw material, use hollow-fibre membrane micro-filtration method, remove foreign protein and DNA in encephalitis B used for human being vaccine harvest liquid, the liquid of collecting after filtering is made encephalitis B used for human being vaccine concentrated solution through ultrafiltration and concentration system, uses sieve chromatography purification system to carry out purification to encephalitis B used for human being vaccine inactivation liquid after deactivation.
This method of utilizing hollow-fibre membrane to remove residual DNA in encephalitis B used for human being vaccine product, its preferred scheme is:
Choose the encephalitis B used for human being vaccine virus harvest liquid of batch, use hollow-fibre membrane to carry out micro-filtration processing to it, hollow-fibre membrane system is that the model that GE company produces is CFP-6-D-8A, and aperture is 0.65 μ m, and membrane area is 4.0m
2liquor inlet end pressure < 10PSI, material liquid outlet end pressure is>=0PSI, feed liquid backflow end pressure < 3PSI, the encephalitis B used for human being vaccine virus harvest liquid of processing, cumulative volume is 230L-240L, and net cycle time is 1.5h-2.0h, guarantees that feed temperature is between 18 ℃-26 ℃ in whole micro-filtration processing procedure.The liquid obtaining after filtration enters into concentration technology program, be made into Vaccinum Encephalitis B concentrated solution by ultrafiltration and concentration system, after deactivation, lot number is SA-03, then by molecular sieve layer analysis system purification, the encephalitis B used for human being vaccine refined solution lot number of collecting is SA-04.
This method of utilizing hollow-fibre membrane to remove residual DNA in encephalitis B used for human being vaccine product, its preferred scheme is:
Choose the encephalitis B used for human being vaccine virus harvest liquid of batch, use hollow-fibre membrane to carry out micro-filtration processing to it, the model of hollow-fibre membrane system is CFP-6-D-8A, and aperture is 0.65 μ m, and membrane area is 4.0m
2liquor inlet end pressure < 10PSI, material liquid outlet end pressure is>=0PSI, feed liquid backflow end pressure < 3PSI, the encephalitis B used for human being vaccine virus harvest liquid of processing, cumulative volume is 110L-120L, and net cycle time is 0.8h-1.0h, guarantees that feed temperature is between 18 ℃-26 ℃ in whole micro-filtration processing procedure.The liquid obtaining after filtration enters into concentration technology program, be made into Vaccinum Encephalitis B concentrated solution by ultrafiltration and concentration system, after deactivation, lot number is SA-05, then by molecular sieve layer analysis system purification, the encephalitis B used for human being vaccine refined solution lot number of collecting is SA-06.
Hollow-fibre membrane has the open flow passage structure of fiber tubulose; avoided the random high turbulences of feed liquid without the tubulose flow passage structure of screen cloth; therefore there is lower shearing force; gentle operation can effectively prevent the gathering of coming off of virus surface glycoprotein and albumen; be conducive to protect the integrity of macromole virus, prevent that virion from contributing to seeing through and removing of foreign protein and DNA when gathering.
Vaccine virus harvest liquid is after filtration treatment, enter into concentration technology program, be made into vaccine virus concentrated solution by ultrafiltration and concentration system, in this process, in the ultrafiltration permeate obtaining, can contain the foreign protein of some and DNA. to vaccine virus harvest liquid, take different filter types, in the ultrafiltration permeate that correspondence obtains, the content of DNA has notable difference.The concrete data that detect are in table 3.
Compared with prior art, the invention has the beneficial effects as follows:
1. hollow-fibre membrane has good physical chemistry toleration, and liquor inlet is withstand voltage is greater than 4bar, can be below 50 degrees Celsius, and the thorough cleaning and sterilizing of tolerance 1mol/L NaOH solution, all right autoclaving of most of hollow-fibre membranes, the life-span is long, and cost is low.
2. hollow-fibre membrane is carrying out, in separation process, the variation of matter not occurring to solution, and chemical compatibility is superior, consumes energy low, and technological process is simple, easy operating management.
Accompanying drawing explanation
Fig. 1 SA-02 purification collection of illustrative plates
Encephalitis B used for human being vaccine inactivation liquid (SA-01), the directly collection of illustrative plates of (SA-02) after sieve chromatography purification, vertical coordinate in figure (ultraviolet absorption value mAU), abscissa (volume ml).
Fig. 2 SA-04 purification collection of illustrative plates
Encephalitis B used for human being vaccine inactivation liquid (SA-03), first use after hollow-fibre membrane micro-filtration, carry out again the collection of illustrative plates of (SA-04) after sieve chromatography purification, vertical coordinate in figure (ultraviolet absorption value mAU, abscissa (volume ml).
Fig. 3 SA-06 purification collection of illustrative plates
Encephalitis B used for human being vaccine inactivation liquid (SA-05), first use after hollow-fibre membrane micro-filtration, carry out again the collection of illustrative plates of (SA-06) after sieve chromatography purification, vertical coordinate in figure (ultraviolet absorption value mAU), abscissa (volume ml).
The specific embodiment
Following examples describe technical scheme of the present invention in detail, limit the scope of the invention but be not used in.
Embodiment 1
Choose the Vaccinum Encephalitis B virus harvest liquid of batch, use sleeve filter to carry out clarification filtration processing to it, the model of sleeve filter system is TP012TV25TCTFELB, and aperture is 0.65 μ m, and membrane area is about 4.0m
2, liquor inlet pressure < 10PSI, material liquid outlet pressure is about 0PSI, the encephalitis B used for human being vaccine harvest liquid of processing, cumulative volume is 230L-240L, net cycle time is 1.5h-2.0h, in whole clarification filtration processing procedure, guarantee that feed temperature is between 18 ℃-26 ℃, the liquid obtaining after filtration enters into concentration technology program, be made into encephalitis B used for human being vaccine concentrated solution by ultrafiltration and concentration system, after deactivation, lot number is SA-01, again by molecular sieve layer analysis system purification, the encephalitis B used for human being vaccine refined solution lot number of collecting is SA-02.
In concentration technology program: ultrafiltration permeate lot number is SC-01.
Embodiment 2
Choose the encephalitis B used for human being vaccine virus harvest liquid of batch, use hollow-fibre membrane to carry out micro-filtration processing to it, the model of hollow-fibre membrane system is CFP-6-D-8A, and aperture is 0.65 μ m, and membrane area is about 4.0m
2liquor inlet end pressure < 10PSI, material liquid outlet end pressure is about 0PSI, feed liquid backflow end pressure < 3PSI, the encephalitis B used for human being vaccine virus harvest liquid of processing, cumulative volume is 230L-240L, and net cycle time is 1.5h-2.0h, guarantees that feed temperature is between 18 ℃-26 ℃ in whole micro-filtration processing procedure.The liquid obtaining after filtration enters into concentration technology program, be made into Vaccinum Encephalitis B concentrated solution by ultrafiltration and concentration system, after deactivation, lot number is SA-03, then by molecular sieve layer analysis system purification, the encephalitis B used for human being vaccine refined solution lot number of collecting is SA-04.
In concentration technology program: ultrafiltration permeate lot number is SC-02.
Embodiment 3
Choose the encephalitis B used for human being vaccine virus harvest liquid of batch, use hollow-fibre membrane to carry out micro-filtration processing to it, the model of hollow-fibre membrane system is CFP-6-D-8A, and aperture is 0.65 μ m, and membrane area is about 4.0m
2liquor inlet end pressure < 10PSI, material liquid outlet end pressure is about 0PSI, feed liquid backflow end pressure < 3PSI, the encephalitis B used for human being vaccine virus harvest liquid of processing, cumulative volume is 110L-120L, and net cycle time is 0.8h-1.0h, guarantees that feed temperature is between 18 ℃-26 ℃ in whole micro-filtration processing procedure.The liquid obtaining after filtration enters into concentration technology program, be made into Vaccinum Encephalitis B concentrated solution by ultrafiltration and concentration system, after deactivation, lot number is SA-05, then by molecular sieve layer analysis system purification, the encephalitis B used for human being vaccine refined solution lot number of collecting is SA-06.
In concentration technology program: ultrafiltration permeate lot number is SC-03.
By SA-01, SA-02, SA-03, SA-04, SA-05, SA-06 respectively through row antigenic content, DNA residues detection.Detect data in table 1.
SC-01, SC-02, SC-03 are carried out respectively to DNA content detection.Detect data and refer to table 3.
Table 1:SA-01～SA-06 antigenic content, DNA residues detection result.
Table 2: different process detects data comparison after processing for identical encephalitis B used for human being viral vaccine concentrated solution
Table 3: in ultrafiltration permeate, DNA content detects data comparison
SC-01 SC-02 SC-03
DNA content 0.8Ng/1ml 1.2Ng/1ml 1.3Ng/1ml
Can be reached a conclusion by table 1, at encephalitis B used for human being vaccine virus antigen, almost in break-even situation, the successful that hollow-fibre membrane system is removed residual DNA in vaccine product is better than sleeve filter filtration system.Hollow-fibre membrane has uniform pore-size distribution, has avoided to a certain extent the accumulation of foreign protein and DNA, is conducive to prevent the formation of film surface protein gel layer, thereby has processing speed faster, is beneficial to and improves later stage chromatography degree.In sum, the unique texture of hollow-fibre membrane, has a certain proportion of removal ability for DNA residual in vaccine product, can reach the object of the more residual DNAs of removal completely.
As shown in Table 3, vaccine virus harvest liquid is after hollow-fibre membrane system micro-filtration is processed, the increased content of DNA in ultrafiltration permeate, obviously in explanation vaccine virus concentrated solution, residual DNA is reducing, fully prove, hollow-fibre membrane system has the removal effect of certain ability for DNA residual in vaccine product, can reach the object of effective removal residual DNA, guarantees that final products reach national acceptable quality.Above-mentioned data detection method all meets the Pharmacopoeia of the People's Republic of China 2010 editions.
Claims (3)
1. a method of utilizing hollow-fibre membrane to remove residual DNA in encephalitis B used for human being vaccine product, it is characterized in that: take encephalitis B used for human being vaccine harvest liquid as raw material, use hollow-fibre membrane micro-filtration method, remove foreign protein and DNA in encephalitis B used for human being vaccine harvest liquid, the liquid of collecting after filtering is made encephalitis B used for human being vaccine concentrated solution through ultrafiltration and concentration system, uses sieve chromatography purification system to carry out purification to encephalitis B used for human being vaccine inactivation liquid after deactivation.
2. according to the method for utilizing hollow-fibre membrane to remove residual DNA in encephalitis B used for human being vaccine product claimed in claim 1, it is characterized in that preferred scheme is:
Choose the encephalitis B used for human being vaccine virus harvest liquid of batch, use hollow-fibre membrane to carry out micro-filtration processing to it, hollow-fibre membrane system is that the model that GE company produces is CFP-6-D-8A, and aperture is 0.65 μ m,, membrane area is 4.0m
2, liquor inlet end pressure < 10PSI, material liquid outlet end pressure is>=0PSI, feed liquid backflow end pressure < 3PSI, the encephalitis B used for human being vaccine virus harvest liquid of processing, cumulative volume is 230L-240L, net cycle time is 1.5h-2.0h, in whole micro-filtration processing procedure, guarantee that feed temperature is between 18 ℃-26 ℃, the liquid obtaining after filtration enters into concentration technology program, be made into Vaccinum Encephalitis B concentrated solution (after deactivation, lot number is SA-03) by ultrafiltration and concentration system, again by molecular sieve layer analysis system purification, that collects is encephalitis B used for human being vaccine refined solution (lot number is SA-04).
3. according to the method for utilizing hollow-fibre membrane to remove residual DNA in encephalitis B used for human being vaccine product claimed in claim 1, it is characterized in that preferred scheme is:
Choose the encephalitis B used for human being vaccine virus harvest liquid of batch, use hollow-fibre membrane to carry out micro-filtration processing to it, the model of hollow-fibre membrane system is CFP-6-D-8A, and aperture is 0.65 μ m, and membrane area is 4.0m
2, liquor inlet end pressure < 10PSI, material liquid outlet end pressure is>=0PSI, feed liquid backflow end pressure < 3PSI, the encephalitis B used for human being vaccine virus harvest liquid of processing, cumulative volume is 110L-120L, net cycle time is 0.8h-1.0h, in whole micro-filtration processing procedure, guarantee that feed temperature is between 18 ℃-26 ℃, the liquid obtaining after filtration enters into concentration technology program, be made into Vaccinum Encephalitis B concentrated solution (after deactivation, lot number is SA-05) by ultrafiltration and concentration system, again by molecular sieve layer analysis system purification, that collects is encephalitis B used for human being vaccine refined solution (lot number is SA-06).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210398655.XA CN103768589B (en) | 2012-10-18 | 2012-10-18 | Hollow-fibre membrane is utilized to remove the method for residual DNA in encephalitis B used for human being vaccine product |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210398655.XA CN103768589B (en) | 2012-10-18 | 2012-10-18 | Hollow-fibre membrane is utilized to remove the method for residual DNA in encephalitis B used for human being vaccine product |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103768589A true CN103768589A (en) | 2014-05-07 |
| CN103768589B CN103768589B (en) | 2016-12-21 |
Family
ID=50561647
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210398655.XA Active CN103768589B (en) | 2012-10-18 | 2012-10-18 | Hollow-fibre membrane is utilized to remove the method for residual DNA in encephalitis B used for human being vaccine product |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103768589B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005080556A2 (en) * | 2004-02-23 | 2005-09-01 | Crucell Holland B.V. | Virus purification methods |
| CN101270350A (en) * | 2008-03-05 | 2008-09-24 | 辽宁依生生物制药有限公司 | Method for extracting rabies virus |
| CN102406933A (en) * | 2011-11-25 | 2012-04-11 | 成都康华生物制品有限公司 | Preparation method for measles attenuated live vaccine |
-
2012
- 2012-10-18 CN CN201210398655.XA patent/CN103768589B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005080556A2 (en) * | 2004-02-23 | 2005-09-01 | Crucell Holland B.V. | Virus purification methods |
| CN101270350A (en) * | 2008-03-05 | 2008-09-24 | 辽宁依生生物制药有限公司 | Method for extracting rabies virus |
| CN102406933A (en) * | 2011-11-25 | 2012-04-11 | 成都康华生物制品有限公司 | Preparation method for measles attenuated live vaccine |
Non-Patent Citations (1)
| Title |
|---|
| 陈昕等: "纯化病毒疫苗的通用方法", 《色谱园》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103768589B (en) | 2016-12-21 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107384877A (en) | A kind of purification process of slow virus | |
| CN107267466B (en) | Method for large-scale production of porcine pseudorabies inactivated vaccine | |
| CN103554253B (en) | A kind of preparation method of quiet note human normal immunoglobulin | |
| CN1204148C (en) | Separation and purification process of lentinan | |
| CN104027800A (en) | Method for preparing rabies vaccines for human use | |
| CN103937754B (en) | Porcine reproductive and respiratory syndrome virus (PPRSV) purification method | |
| CN115141813A (en) | Adenovirus purification method for efficiently removing residual proteins of host cells | |
| CN103768589A (en) | Method for removing residual DNA in encephalitis B vaccine product by utilizing hollow fiber membrane | |
| CN104450656A (en) | Method for preparing and purifying thrombin in rabbit blood | |
| JP2000317273A (en) | Membrane separation method | |
| CN106279376A (en) | A kind of pig circular ring virus antigen purification method | |
| CN101380467B (en) | Multi-connection inactivated vaccine (antigen) liquid concentration technique | |
| CN109134622A (en) | The multistep continuous Integration purification process of foot-and-mouth disease virus antigen | |
| CN103768590A (en) | Method for removing residual DNA in encephalitis B vaccine product by utilizing anionic exchange chromatography | |
| CN101780134B (en) | Membrane separation process for extracting solution of south dodder seed | |
| CN108606156A (en) | A kind of wheat oligopeptide and its industrialized preparing process | |
| CN103480276B (en) | A kind of closed ultrafiltration pipe-line system and application thereof | |
| CN114525263A (en) | Purification and concentration method of porcine acute diarrhea syndrome coronavirus antigen | |
| CN207628222U (en) | Spent solvent system processing system | |
| CN105200038A (en) | Scale purification method for plasmid DNA | |
| CN106188338B (en) | A method of reducing foreign protein content in hyaluronic acid sodium raw materials | |
| CN104017037B (en) | A kind of preparation method of the former powder of jinggangmeisu | |
| CN203447991U (en) | Closed type ultra-filtration pipeline system | |
| CN208414394U (en) | A kind of enrichment facility of microorganism sewage water processing strain | |
| CN109651470A (en) | A kind of separation method of Coenzyme I and other macromolecular substances |
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 |