DK1869171T4 - Virusrensning under anvendelse af ultrafiltrering - Google Patents
Virusrensning under anvendelse af ultrafiltrering Download PDFInfo
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- DK1869171T4 DK1869171T4 DK06724513.4T DK06724513T DK1869171T4 DK 1869171 T4 DK1869171 T4 DK 1869171T4 DK 06724513 T DK06724513 T DK 06724513T DK 1869171 T4 DK1869171 T4 DK 1869171T4
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- Prior art keywords
- adenovirus
- virus
- buffer
- cells
- back pressure
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
- C12N2710/00011—Details
- C12N2710/10011—Adenoviridae
- C12N2710/10051—Methods of production or purification of viral material
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- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Virology (AREA)
- Biotechnology (AREA)
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- Medicinal Chemistry (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Immunology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Separation Using Semi-Permeable Membranes (AREA)
Claims (20)
1. Fremgangsmåde til rensning af et rekombinant adenovirus, omfattende et ultrafiltreringstrin, hvor retentatet indeholder viruset, kendetegnet ved, at et modtryk på mindst 5 kPa påsættes på permeatsiden.
2. Fremgangsmåde ifølge krav 1, hvor fremgangsmåden inden ultrafiltreringstrinnet omfatter følgende trin: a) dyrkning af celler, der er inficeret med viruset, b) tilsætning af nuklease til cellekulturen og derefter c) eventuelt lyse af cellerne til tilvejebringelse af et lysat indeholdende viruset og d) eventuelt klaring af lysatet, fortrinsvis ved dybfiltrering efterfulgt af membranfiltrering.
3. Fremgangsmåde ifølge ethvert af de foregående krav, ved hvilken ultrafiltreringen omfatter tangentiel strømningsfiltrering, fortrinsvis under anvendelse af et hult fibermodul.
4. Fremgangsmåde ifølge ethvert af de foregående krav, ved hvilken modtrykket på mindst 5 kPa påsættes med en pumpe, der tilvejebringer modtryk til permeatet.
5. Fremgangsmåde ifølge ethvert af de foregående krav, ved hvilken transmembrantrykket holdes under 20 kPa.
6. Fremgangsmåde ifølge ethvert af de foregående krav, ved hvilken ultrafiltreringen omfatter bufferskift i retentatet med en buffer indeholdende mellem 0,8 M og 2 M natriumchlorid eller et andet salt, der giver samme ionstyrke, og fortrinsvis bufferskift med en buffer med en ionstyrke som en buffer indeholdende mindre end 0,5 M NaCI.
7. Fremgangsmåde ifølge ethvert af de foregående krav, hvilken fremgangsmåde ikke omfatter et trin til størrelsesudelukkelseskromatografi.
8. Fremgangsmåde ifølge ethvert af de foregående krav, hvilken fremgangsmåde ikke omfatter et trin til anionbytningskromatografi eller anionbytningsfiltrering.
9. Fremgangsmåde ifølge ethvert af de foregående krav, hvilken fremgangsmåde er kendetegnet ved, at den ikke omfatter et kromatografitrin eller et ionbytningsfiltreringstrin.
10. Fremgangsmåde ifølge ethvert af de foregående krav, hvilken fremgangsmåde endvidere omfatter trin til yderligere rensning af det rekombinante adenovirus med mindst ét kromatografitrin.
11. Fremgangsmåde ifølge krav 10, ved hvilken fremgangsmåden omfatter et trin til anionbytningskromatografi, eventuelt udført ved påføring af den virusholdige opløsning på et ladet filter, der indeholder positivt ladede grupper.
12. Fremgangsmåde ifølge krav 10 eller 11, hvor fremgangsmåden omfatter et trin til størrelsesudelukkelseskromatografi.
13. Fremgangsmåde til rensning af et rekombinant adenovirus, hvilken fremgangsmåde i det væsentlige består af: a) dyrkning af celler, der er inficeret med det rekombinante adenovirus, b) lyse af cellerne og fjernelse og/eller fragmentering af fri nukleinsyre til tilvejebringelse af et lysat indeholdende det rekombinante adenovirus, c) klaring af lysatet til opnåelse af et adenoviruspræparat, d) udsættelse af adenoviruspræparatet for ultrafiltrering, hvor adenoviruspræparatet befinder sig i retentatet, til koncentrering af adenoviruspræparatet, e) udsættelse af adenoviruspræparatet fra trin d) for ultrafiltrering, hvor adenoviruspræparatet befinder sig i retentatet, og udskiftning deraf med mindst 5 diafiltreringsrumfang (DFV’er) buffer, hvor én DFV er rumfanget af retentatet efter koncentrering i trin d), f) fortrinsvis sterilfiltrering af adenoviruspræparat, hvilken fremgangsmåde er kendetegnet ved, at der i trin d) og e) påsættes et modtryk på mindst 5 kPa på permeatsiden.
14. Fremgangsmåde til rensning af et rekombinant adenovirus, hvilken fremgangsmåde i det væsentlige består af: a) dyrkning af celler, der er inficeret med det rekombinante adenovirus, b) lyse af cellerne og fjernelse og/eller fragmentering af fri nukleinsyre til tilvejebringelse af et lysat indeholdende det rekombinante adenovirus, c) klaring af lysatet til opnåelse af et adenoviruspræparat, d) udsættelse af adenoviruspræparatet fra trin c) for ultrafiltrering, hvor adenovirus-præparatet befinder sig i retentatet, og udskiftning deraf med mindst 5 diafiltreringsrumfang (DFV’er) buffer, hvor én DFV er rumfanget af adenoviruspræparatet efter c), e) fortrinsvis sterilfiltrering af adenoviruspræparatet, hvilken fremgangsmåde er kendetegnet ved, at i trin d) påsættes et modtryk på mindst 5 kPa på permeatsiden.
15. Fremgangsmåde ifølge krav 13 eller 14, ved hvilken trin b) omfatter: b, i) tilsætning af nuklease til cellekulturen og derefter b, ii) lyse af cellerne til tilvejebringelse af et lysat indeholdende det rekombinante adenovirus.
16. Fremgangsmåde ifølge ethvert af kravene 13-15, ved hvilken ultrafiltreringen omfatter tangentiel strømningsfiltrering, fortrinsvis under anvendelse af et hult fibermodul.
17. Fremgangsmåde ifølge ethvert af kravene 13-16, ved hvilken modtrykket påsættes med en pumpe, der tilvejebringer modtryk til permeatet.
18. Fremgangsmåde ifølge ethvert af kravene 13-17, ved hvilken transmembrantrykket under ultrafiltreringen holdes under 20 kPa.
19. Fremgangsmåde ifølge ethvert af kravene 13-18, ved hvilken bufferskiftet med retentatet omfatter bufferskift med buffere med forskellig ionstyrke, herunder bufferskift med en buffer indeholdende mellem 0,8 M og 2 M natriumchlorid eller et andet salt, der giver samme ionstyrke, og fortrinsvis efterfølgende bufferskift med en buffer med en ionstyrke som en buffer indeholdende mindre end 0,5 M NaCI.
20. Fremgangsmåde til forøgelse af udvindingen og/eller udbyttet af rekombinant adenovirus under et ultrafiltreringstrin, hvor retentatet indeholder det rekombinante adenovirus, hvilken fremgangsmåde er kendetegnet ved, at et modtryk på mindst 5 kPa påsættes på permeatsiden.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US67006405P | 2005-04-11 | 2005-04-11 | |
| EP05102842 | 2005-04-11 | ||
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| US (1) | US8574595B2 (da) |
| EP (1) | EP1869171B2 (da) |
| CN (1) | CN101155915B (da) |
| AT (1) | ATE412737T1 (da) |
| AU (1) | AU2006233800B2 (da) |
| CA (1) | CA2602944C (da) |
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| DK (1) | DK1869171T4 (da) |
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2006
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Also Published As
| Publication number | Publication date |
|---|---|
| EP1869171B1 (en) | 2008-10-29 |
| EP1869171A1 (en) | 2007-12-26 |
| DE602006003420D1 (de) | 2008-12-11 |
| AU2006233800A1 (en) | 2006-10-19 |
| CA2602944A1 (en) | 2006-10-19 |
| CN101155915B (zh) | 2013-12-04 |
| US8574595B2 (en) | 2013-11-05 |
| US20090123989A1 (en) | 2009-05-14 |
| ATE412737T1 (de) | 2008-11-15 |
| EP1869171B2 (en) | 2015-10-14 |
| AU2006233800B2 (en) | 2011-07-07 |
| ES2317517T5 (es) | 2016-01-21 |
| DK1869171T3 (da) | 2009-02-16 |
| CN101155915A (zh) | 2008-04-02 |
| ES2317517T3 (es) | 2009-04-16 |
| WO2006108707A1 (en) | 2006-10-19 |
| CA2602944C (en) | 2015-08-11 |
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