DK2675476T3 - Fremgangsmåde til fremstilling af PRRSV i kommerciel skala - Google Patents

Fremgangsmåde til fremstilling af PRRSV i kommerciel skala Download PDF

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DK2675476T3
DK2675476T3 DK12706508.4T DK12706508T DK2675476T3 DK 2675476 T3 DK2675476 T3 DK 2675476T3 DK 12706508 T DK12706508 T DK 12706508T DK 2675476 T3 DK2675476 T3 DK 2675476T3
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virus
harvest
prrsv
bioreactor
medium
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Elizabeth Jane Berry
Fuad Tawfiq Haddadin
Ali Khazraeinazmpour
Jeremy Kroll
Sonia Regina Cantisano Malburg
Basurto Edgar Arnulfo Sandoval
Stephen Scheerer
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Boehringer Ingelheim Vetmed
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    • C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
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    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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    • A—HUMAN NECESSITIES
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    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K39/00—Medicinal preparations containing antigens or antibodies
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    • C12N2770/00011—Details
    • C12N2770/10011—Arteriviridae
    • C12N2770/10021—Viruses as such, e.g. new isolates, mutants or their genomic sequences
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    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
    • C12N2770/00011—Details
    • C12N2770/10011—Arteriviridae
    • C12N2770/10034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
    • C—CHEMISTRY; METALLURGY
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    • C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
    • C12N2770/00011—Details
    • C12N2770/10011—Arteriviridae
    • C12N2770/10051—Methods of production or purification of viral material

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Claims (16)

1. Fremgangsmåde til fremstilling af porcin reproduktions- og respirationssygdomsvirus (PRRSV) i kommerciel skala, hvilken fremgangsmåde omfatter: a) samtidigt at pode et storskalakulturmedium med en pattedyrcellelinje, som tillader PRRSV-infektion, i en bioreaktor og inficere pattedyrcellerne med en PRRSV; b) at opformere virus i 5 til 7 dage efter infektion; c) at gennemføre et første indhøstningstrin ved at fjerne mediet fra bioreaktoren og isolere opformeret virus derfra; d) at genopfylde mediet i bioreaktoren og opformere virus i 1 til 4 dage; e) at gennemføre et andet indhøstningstrin ved at fjerne mediet fra bioreaktoren og isolere opformeret virus derfra; f) at genopfylde mediet i bioreaktoren og opformere virus i 1 til 4 dage, og g) at gennemføre et tredje indhøstningstrin ved at fjerne mediet fra bioreaktoren og isolere opformeret virus derfra.
2. Fremgangsmåde ifølge krav 1, hvilken fremgangsmåde yderligere omfatter mindst et gentilførsels- og indhøstningstrin efter det tredje indhøstningstrin, omfattende at genopfylde medium i bioreaktoren og at opformere virus i 1 til 4 dage og at gennemføre et fjerde indhøstningstrin ved at fjerne mediet fra bioreaktoren og isolere opformeret virus derfra.
3. Fremgangsmåde ifølge krav 1, hvor den tilsigtede infektionsfaktor (MOI) er 0,01 til 0,30.
4. Fremgangsmåde ifølge krav 1, hvor pattedyrcellerne indplantes med en tæthed på cirka 7 x 108 til 1,0 x 109 per 300 I bioreaktor.
5. Fremgangsmåde ifølge krav 4, hvor celleindplantningstætheden er cirka 1,00 x 109 per 300 I bioreaktor.
6. Fremgangsmåde ifølge krav 5, hvor infektionen sker med cirka 7 x 108 viruspartikler per 300 I bioreaktor.
7. Fremgangsmåde ifølge krav 1, omfattende at overvåge dextrosekoncentrationen i mediet, hvor det første indhøstningstrin gennemføres på den første dag, når dextrosekoncentrationen i mediet falder til under 0,1 g/l.
8. Fremgangsmåde ifølge krav 1, hvor den anden indhøstning gennemføres 1 eller 2 dage efter mediegentilførsel.
9. Fremgangsmåde ifølge krav 1, hvor den tredje indhøstning gennemføres 1 til 4 dage efter anden mediegentilførsel.
10. Fremgangsmåde ifølge krav 1, hvor kulturmediet tilsættes til bioreaktoren dagen inden eller same dag forud for indbringning af pattedyrcellelinjen og nævnte PRRS.
11. Fremgangsmåde ifølge krav 1, hvor kulturmediet tilsættes til bioreaktoren én dag inden indbringning af pattedyrcellelinjen og nævnte PRRS.
12. Fremgangsmåde ifølge krav 1, hvor bioreaktorens temperatur indstilles til mellem 34 °C og 38 °C.
13. Fremgangsmåde ifølge krav 1, hvor mediet omfatter 5 % v/v bestrålet kalvefo-sterserum.
14. Fremgangsmåde ifølge krav 1, hvor nævnte PRRSV er udvalgt fra gruppen bestående af PRRSV-stamme ECACC 11012501, ECACC 11012502, VR 2332, Lelystad-virusstamme (Lelystad Agent (CDI-NL-2.91), eller andre stammer såsom hine deponeret under tilgangsnumrene ECACC 04102703, ECACC 04102702, ECACC 04102704, CNCM-tilgangsnr. 1-1140, CNCM-tilgangsnr. 1-1387, CNCM-tilgangsnr. 1-1388, ATCC VR 2332, VR 2385, VR 2386, VR 2429, VR 2474 og VR 2402; CNCM 1-1102, CNCM 1-1140, CNCM I-1387, CNCM 1-1388 eller ECACC V93070108; ATCC-deponering VR-2332, ATCC-deponering VR-2368; ATCC VR-2495; ATCC VR 2385, ATCC VR 2386, ATCC VR 2429, ATCC VR 2474 og ATCC VR 2402.
15. Fremgangsmåde til fremstilling af en PRRSV i kommerciel skala, hvilken fremgangsmåde omfatter: a. samtidig at pode både pattedyrceller, som tillader infektion med nævnte PRRSV, og nævnte PRRSV i en bioreaktor indeholdende et medium, der egner sig til dyrkning af cellerne; og b. at gennemføre tre på hinanden følgende indhøstningstrin, hvor PRRSV indhøstes, idet mediet genopfyldes efter hver af den første og den anden indhøstning, og hvor: i. den første indhøstning gennemføres på den første dag, hvor mediets dextrosekoncentration falder til under 0,1 g/l; ii. den anden indhøstning gennemføres 1 eller 2 dage efter tilsætning af mediet efter den første indhøstning; og iii. den tredje indhøstning gennemføres mellem 1 og 4 dage efter tilsætning af mediet efter den anden indhøstning.
16. Fremgangsmåde til fremstilling i kommerciel skala af porcin reproduktions- og respirationssygdomsvirus (PRRSV), hvilken fremgangsmåde omfatter: a) samtidigt at pode et storskalakulturmedium med en pattedyrcellelinje, som tillader PRRSV-infektion, i en rulleflaske og at inficere pattedyrcellerne med en PRRSV; b) at opformere virus i 5 til 7 dage efter infektion; c) at gennemføre et første indhøstningstrin ved at fjerne mediet fra rulleflasken og isolere opformeret virus derfra; d) at genopfylde mediet i rulleflasken og opformere virus i cirka 2 dage; e) at gennemføre et andet indhøstningstrin ved at fjerne mediet fra rulleflasken og isolere opformeret virus derfra; f) at genopfylde mediet i rulleflasken og opformere virus i cirka 2 dage, og g) at gennemføre et tredje indhøstningstrin ved at fjerne mediet fra bioreaktoren og isolere opformeret virus derfra.
DK12706508.4T 2011-02-17 2012-02-14 Fremgangsmåde til fremstilling af PRRSV i kommerciel skala DK2675476T3 (da)

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US201161444071P 2011-02-17 2011-02-17
PCT/EP2012/052476 WO2012110490A1 (en) 2011-02-17 2012-02-14 Commercial scale process for production of prrsv

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