DK2530161T3 - Fremgangsmåde til fremstilling af poxvira og poxvirussammensætninger - Google Patents

Fremgangsmåde til fremstilling af poxvira og poxvirussammensætninger Download PDF

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DK2530161T3
DK2530161T3 DK12179919.1T DK12179919T DK2530161T3 DK 2530161 T3 DK2530161 T3 DK 2530161T3 DK 12179919 T DK12179919 T DK 12179919T DK 2530161 T3 DK2530161 T3 DK 2530161T3
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virus
cell
recombinant
cells
mva
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Daniel Malarme
Yves Cordier
Claude Sene
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Transgene Sa
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    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C12N15/86—Viral vectors
    • C12N15/863—Poxviral vectors, e.g. entomopoxvirus
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    • A61K2039/58—Medicinal preparations containing antigens or antibodies raising an immune response against a target which is not the antigen used for immunisation
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    • C12N2710/00011—Details
    • C12N2710/20011—Papillomaviridae
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    • C12N2710/24011—Poxviridae
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Claims (14)

1. Fremgangsmåde til fremstilling af en vildtype-, en svækket og/eller en re-kombinant poxvirus med ingen targeted infektionsspecificitet, omfattende trinnene: a) at fremstille en kultur af packaging-celler, b) at inficere cellekulturen, c) at dyrke de inficerede celler i et passende tidsrum, især mellem to og fire dage, d) at genindvinde poxviruspartiklerne produceret fra kultursupernatanten og packaging-cellerne, hvor trin d) omfatter at bryde packaging-cellen op ved anvendelse en højhastigheds-homogenisator, e) et klaringstrin med blandingen opnået fra trin d), som muliggør fjernelsen af celleresterne, hvor klaringstrinnet er et dybdefiltreringstrin, f) et koncentrationstrin, hvor koncentrationstrinnet er et mikrofiltreringstrin, og g) et diafiltreringstrin, hvor der ikke anvendes nuklease og især ikke benzonase, og hvor fremgangsmåden er fri for animalske produkter.
2. Fremgangsmåde ifølge krav 1, hvor packaging-cellen er en primær celle, en rekombinant celle og/eller en cellelinje.
3. Fremgangsmåde ifølge krav 2, hvor packaging-cellen er en CEF.
4. Fremgangsmåde ifølge et af kravene 1 til 3, hvor den svækkede og/eller rekombinante poxvirus med ingen targeted infektionsspecificitet er en rekombinant modificeret vaccinia-virus Ankara (MVA)-virus.
5. Fremgangsmåde ifølge krav 4, hvor den rekombinante MVA-virus omfatter en eksogen sekvens, der koder et molekyle, som har en direkte eller indirekte cytotoksisk funktion, især hvor den eksogene sekvens er et selvmordsgen, der koder et protein med en cytosindeaminase-aktivitet.
6. Fremgangsmåde ifølge krav 4, hvor den rekombinante MVA-virus omfatter en eksogen sekvens, som koder et tumorassocieret antigen (TAA).
7. Fremgangsmåde ifølge krav 6, hvor den rekombinante MVA-virus yderligere omfatter en eksogen sekvens, der koder et cytokin.
8. Fremgangsmåde ifølge krav 6 eller krav 7, hvor TAA er MUC-1.
9. Fremgangsmåde ifølge krav 8, hvor den rekombinante MVA-virus er en MVA-vektor, der bærer kodningssekvenserne for MUC1 -protein under kontrollen af promoter p7.5 og IL2-protein under kontrollen af promoter pFI5R.
10. Fremgangsmåde ifølge krav 4, hvor den rekombinante MVA-virus omfatter en eksogen sekvens, der koder et antigen, især hvor antigenet stammer fra en virus.
11. Fremgangsmåde ifølge krav 10, hvor antigenet er udvalgt blandt: • et non-strukturelt protein fra genotype 1 b stamme ja af hepatitis C-virus, og • E6- og E7-proteiner fra human papillomavirus (FIPV) 16.
12. Fremgangsmåde ifølge krav 11, hvor den rekombinante MVA-virus er en MVA-vektor, der bærer kodningssekvenserne for FIPV-proteiner E6 og E7 under kontrollen af promoter p7.5 og IL2-protein under kontrollen af promoter pH5R.
13. Fremgangsmåde ifølge et af kravene 1 til 12, hvor dybdefiltreringstrin e) forbinder to eller flere enheder med aftagende porestørrelser.
14. Fremgangsmåde ifølge et af kravene 1 til 13, hvor filtreringsmembranerne anvendt i mikrofiltreringstrin f) og/eller i diafiltreringstrin g) har en porestørrelse på mellem 0,01 og 0,15 pm, og mere foretrukket ca. 0,1 pm.
DK12179919.1T 2006-06-20 2007-06-15 Fremgangsmåde til fremstilling af poxvira og poxvirussammensætninger DK2530161T3 (da)

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Application Number Priority Date Filing Date Title
EP06360027 2006-06-20
US86145206P 2006-11-29 2006-11-29
EP07764674.3A EP2029756B1 (en) 2006-06-20 2007-06-15 Process for producing poxviruses and poxvirus compositions

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DK07764674.3T DK2029756T3 (da) 2006-06-20 2007-06-15 Fremgangsmåde til fremstilling af poxvira og poxvirussammensætninger
DK12179919.1T DK2530161T3 (da) 2006-06-20 2007-06-15 Fremgangsmåde til fremstilling af poxvira og poxvirussammensætninger

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US (5) US8058049B2 (da)
EP (2) EP2530161B1 (da)
JP (1) JP5456464B2 (da)
KR (1) KR101082296B1 (da)
CN (1) CN101460627B (da)
AU (1) AU2007263280B2 (da)
BR (1) BRPI0712158B8 (da)
CA (1) CA2656259C (da)
CO (1) CO6141479A2 (da)
CR (1) CR10528A (da)
DK (2) DK2029756T3 (da)
ES (2) ES2671230T3 (da)
HK (1) HK1125405A1 (da)
HU (2) HUE030533T2 (da)
IL (1) IL194978A (da)
MA (1) MA30580B1 (da)
MX (2) MX348525B (da)
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RU (3) RU2489486C2 (da)
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ZA (1) ZA200810658B (da)

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