DK1866342T3 - Separation af forurenende kontaminanter fra streptococcus pneumoniae-polysaccharid ved ph-manipulation - Google Patents

Separation af forurenende kontaminanter fra streptococcus pneumoniae-polysaccharid ved ph-manipulation Download PDF

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DK1866342T3
DK1866342T3 DK06740302.2T DK06740302T DK1866342T3 DK 1866342 T3 DK1866342 T3 DK 1866342T3 DK 06740302 T DK06740302 T DK 06740302T DK 1866342 T3 DK1866342 T3 DK 1866342T3
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polysaccharide
precipitate
solution
lysate
detergent
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DK06740302.2T
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Brian Bahler
Tsu-Shun Lee
Jason Arnold Lotvin
Mark Edward Ruppen
Pamela Sue Fink Charbonneau
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Wyeth Llc
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    • CCHEMISTRY; METALLURGY
    • C08ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08BPOLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00Medicinal preparations containing antigens or antibodies
    • A61K39/02Bacterial antigens
    • A61K39/085Staphylococcus
    • CCHEMISTRY; METALLURGY
    • C08ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08BPOLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/0003General processes for their isolation or fractionation, e.g. purification or extraction from biomass
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/06Lysis of microorganisms
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20Bacteria; Culture media therefor
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P1/00Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
    • C12P1/04Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using bacteria
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00Preparation of compounds containing saccharide radicals
    • C12P19/04Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
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  • Tropical Medicine & Parasitology (AREA)
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  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Peptides Or Proteins (AREA)

Claims (8)

1. Fremgangsmåde til at reducere proteinindholdet og opretholde kapselpo-lysaccharid-indholdet i en kompleks cellulær Streptococcus pneumoniae-lysatsuppe før oprensning, hvor fremgangsmåden omfatter følgende trin: (a) at udvide en udvalgt S. pneumoniae-serotype i stigende voluminer fra en begyndelsesbeholder til en beholder i produktionsskala i et sojabaseret medium og opretholde en stabil pH-værdi og temperatur under cellevækst; (b) at lysere bakteriecellerne produceret i trin (a) med en detergent, hvorved et lysat produceres, der indeholder opløselige proteiner, celledebris, nukleinsyrer og polysaccharid; (c) at omrøre det cellulære lysat i et tidsrum, der er tilstrækkeligt til at sikre komplet lysis- og polysaccharidfrigivelse; (d) at reducere pH-værdien af det cellulære lysat til mindre end 5,5 for at udskille detergentet og det meste af de opløselige proteiner; (e) at holde opløsningen og præcipitatet dannet i trin (d) uden omrøring i et tidsrum, der er tilstrækkeligt til at tillade bundfældning af præcipitatet, og (f) at behandle opløsningen og præcipitatet ved centrifugering og/eller filtrering, hvorved kapselpolysaccharidet i opløsningen fastholdes, og det opløselige protein reduceres effektivt.
2. Fremgangsmåde ifølge krav 1, hvor den udvalgte S. pneumoniae-serotype er 1,4, 5, 6A, 6B, 7F eller 19A.
3. Fremgangsmåde ifølge krav 1, hvor pH-værdien i trin (a) opretholdes afen basetilførsel af natriumhydroxid, natriumcarbonat eller en kombination deraf.
4. Fremgangsmåde ifølge krav 1, hvor fremgangsmåden omfatter trinnene: (a) at udvide i stigende voluminer fra en begyndelsesbeholder til en beholder i produktionsskala i et sojabaseret medium et S. pneumoniae-serotype udvalgt fra gruppen bestående af serotyper 1,4, 6A, 6B og 7F og opretholde en stabil pH-værdi og temperatur under cellevækst, hvor pH-værdien opretholdes med natriumhydroxid; (b) at lysere bakteriecellerne produceret i trin (a) med en detergent, hvorved et lysat produceres, der indeholder opløselige proteiner, celledebris, nukleinsyrer og polysaccharid; (c) at omrøre det cellulære lysat i et tidsrum, der er tilstrækkeligt til at sikre komplet lysis- og polysaccharidfrigivelse; (d) at reducere pH-værdien af det cellulære lysat til mindre end 5,5 for at udskille detergentet og det meste af de opløselige proteiner; (e) at holde opløsningen og præcipitatet dannet i trin (d) uden omrøring i et tidsrum, der er tilstrækkeligt til at tillade bundfældning af præcipitatet; og (f) at behandle opløsningen og præcipitatet ved centrifugering og/eller filtrering, hvorved kapselpolysaccharidet i opløsningen fastholdes, og det opløselige protein reduceres effektivt.
5. Fremgangsmåde ifølge krav 1, hvor fremgangsmåden omfatter trinnene: (a) at udvide i stigende voluminer S. pneumoniae-serotype 5 fra en begyndelsesbeholder til en beholder i produktionsskala i et sojabaseret medium suppleret med natriumbikarbonat og opretholde en stabil pH-værdi og temperatur under cellevækst, hvor pH-værdien opretholdes med natriumhydroxid; (b) at lysere bakteriecellerne produceret i trin (a) med en detergent, hvorved et lysat produceres, der indeholder opløselige proteiner, celledebris, nukleinsyrer og polysaccharid; (c) at omrøre det cellulære lysat i et tidsrum, der er tilstrækkeligt til at sikre komplet lysis- og polysaccharidfrigivelse; (d) reducere pH-værdien for det cellulære lysat til mindre end 5,5 for at udfælde detergentet og det meste af de opløselige proteiner; (e) at holde opløsningen og præcipitatet dannet i trin (d) uden omrøring i et tidsrum, der er tilstrækkeligt til at tillade bundfældning af præcipitatet; og (f) at behandle opløsningen og præcipitatet ved centrifugering og/eller filtrering, hvorved kapselpolysaccharidet i opløsningen fastholdes, og det opløselige protein reduceres effektivt.
6. Fremgangsmåde ifølge krav 1, hvor fremgangsmåden omfatter trinnene: (a) at udvide i stigende voluminer S. pneumoniae serotype 19Afra en begyndelsesbeholder til en beholder i produktionsskala i et sojabaseret medium suppleret med natriumbikarbonat og opretholde en stabil pH-værdi og temperatur under cellevækst, hvor pH-værdien opretholdes med natriumcarbonat; (b) at lysere bakteriecellerne produceret i trin (a) med en detergent, hvorved et lysat produceres, der indeholder opløselige proteiner, celledebris, nukleinsyrer og polysaccharid; (c) at omrøre det cellulære lysat i et tidsrum, der er tilstrækkeligt til at sikre komplet lysis- og polysaccharidfrigivelse; (d) at reducere pH-værdien af det cellulære lysat til mindre end 5,5 for at udskille detergentet og det meste af de opløselige proteiner; (e) at holde opløsningen og præcipitatet dannet i trin (d) uden omrøring i et tidsrum, der er tilstrækkeligt til at tillade bundfældning af præcipitatet, og (f) at behandle opløsningen og præcipitatet ved centrifugering og/eller filtrering, hvorved kapselpolysaccharidet i opløsningen fastholdes, og det opløselige protein reduceres effektivt.
7. Fremgangsmåde ifølge et af kravene 1,4,5 eller 6, hvor detergentet er de-oxycholatnatrium.
8. Fremgangsmåde ifølge et af kravene 1, 4, 5 eller 6, hvor pH-værdien i trin (d) reduceres til mellem 4,5 og mindre end 5,5.
DK06740302.2T 2005-04-08 2006-03-31 Separation af forurenende kontaminanter fra streptococcus pneumoniae-polysaccharid ved ph-manipulation DK1866342T3 (da)

Applications Claiming Priority (2)

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US66954605P 2005-04-08 2005-04-08
PCT/US2006/012134 WO2006110352A2 (en) 2005-04-08 2006-03-31 Separation of contaminants from streptococcus pneumoniae polysaccharide by ph manipulation

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US (1) US7718791B2 (da)
EP (2) EP1866342B1 (da)
JP (1) JP5049264B2 (da)
KR (1) KR101317256B1 (da)
CN (1) CN101155833B (da)
AR (1) AR053843A1 (da)
AU (1) AU2006234984B2 (da)
BR (1) BRPI0607026B1 (da)
CA (1) CA2604362C (da)
DK (2) DK3466982T3 (da)
ES (2) ES2707202T3 (da)
HU (1) HUE050049T2 (da)
IL (1) IL186366A (da)
MX (1) MX2007012337A (da)
NZ (1) NZ562719A (da)
PL (2) PL1866342T3 (da)
PT (2) PT3466982T (da)
SI (2) SI3466982T1 (da)
TR (1) TR201820018T4 (da)
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