WO1998022588A2 - An improved method for the production and purification of adenoviral vectors - Google Patents

An improved method for the production and purification of adenoviral vectors Download PDF

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Publication number
WO1998022588A2
WO1998022588A2 PCT/US1997/021504 US9721504W WO9822588A2 WO 1998022588 A2 WO1998022588 A2 WO 1998022588A2 US 9721504 W US9721504 W US 9721504W WO 9822588 A2 WO9822588 A2 WO 9822588A2
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Prior art keywords
cells
cell
adenovims
antisense
vims
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PCT/US1997/021504
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English (en)
French (fr)
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WO1998022588A3 (en
WO1998022588A9 (en
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Shuyuan Zhang
Capucine Thwin
Zheng Wu
Toohyon Cho
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Introgen Therapeutics Inc
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Introgen Therapeutics Inc
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Priority to BR9713368-0A priority Critical patent/BR9713368A/pt
Priority to DE69737107T priority patent/DE69737107T2/de
Priority to EP97950677A priority patent/EP0968284B1/en
Priority to AU53617/98A priority patent/AU732703B2/en
Priority to JP52396598A priority patent/JP4492826B2/ja
Application filed by Introgen Therapeutics Inc filed Critical Introgen Therapeutics Inc
Priority to CA2272820A priority patent/CA2272820C/en
Priority to CN971812543A priority patent/CN1244215B/zh
Publication of WO1998022588A2 publication Critical patent/WO1998022588A2/en
Publication of WO1998022588A3 publication Critical patent/WO1998022588A3/en
Publication of WO1998022588A9 publication Critical patent/WO1998022588A9/en
Priority to NO992389A priority patent/NO992389L/no
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C12N15/86—Viral vectors
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/76—Viruses; Subviral particles; Bacteriophages
    • A61K35/761—Adenovirus
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
    • C07K14/4746—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used p53
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • 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
    • C12N2710/10041—Use of virus, viral particle or viral elements as a vector
    • C12N2710/10045—Special targeting system for viral vectors
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • 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
    • C12N2710/10311—Mastadenovirus, e.g. human or simian adenoviruses
    • C12N2710/10322—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
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    • 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
    • C12N2710/10311—Mastadenovirus, e.g. human or simian adenoviruses
    • C12N2710/10332—Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent
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    • C12N2710/00011—Details
    • C12N2710/10011—Adenoviridae
    • C12N2710/10311—Mastadenovirus, e.g. human or simian adenoviruses
    • C12N2710/10341—Use of virus, viral particle or viral elements as a vector
    • C12N2710/10343—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
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    • 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
    • C12N2710/10311—Mastadenovirus, e.g. human or simian adenoviruses
    • C12N2710/10351—Methods of production or purification of viral material
    • C—CHEMISTRY; METALLURGY
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    • C12N2810/00—Vectors comprising a targeting moiety
    • C12N2810/50—Vectors comprising as targeting moiety peptide derived from defined protein
    • C12N2810/60—Vectors comprising as targeting moiety peptide derived from defined protein from viruses
    • C12N2810/6009—Vectors comprising as targeting moiety peptide derived from defined protein from viruses dsDNA viruses
    • C12N2810/6018—Adenoviridae

Definitions

  • the method further comprises isolating an adenoviral particle from the lysate using chromatography.
  • the isolating consists essentially of a single chromatography step.
  • the chromatography step is ion exchange chromatography.
  • the ion exchange chromatography is carried out at a pH range of between about 7.0 and about 10.0.
  • the ion exchange chromatography is anion exchange chromatography.
  • the present invention involves a process that has been developed for the production and purification of a replication deficient recombinant adenovirus.
  • the production process is based on the use of a CellcubeTM bioreactor for cell growth and virus production. It was found that a given perfusion rate, used during cell growth and the virus production phases of culturing, has a significant effect on the downstream purification of the virus. More specifically, a low to medium perfusion rate improves virus production.
  • lysis solution composed of buffered detergent, used to lyse cells in the CellcubeTM at the end of virus production phase also improves the process. With these two advantages, the harvested crude virus solution can be purified using a single ion exchange chromatography run, after concentration/diafiltration and nuclease treatment to reduce the contaminating nucleic
  • the ability to produce infectious viral vectors is increasingly important to the pharmaceutical industry, especially in the context of gene therapy. Over the last decade, advances in biotechnology have led to the production of a number of important viral vectors that have potential uses as therapies, vaccines and protein production machines.
  • the use of viral vectors in mammalian cultures has advantages over proteins produced in bacterial or other lower lifeform hosts in their ability to post-translationally process complex protein structures such as disulfide-dependent folding and glycosylation.
  • the former can be anionic such as sodium dodecyl sulfate or cationic such as ethyl trimethyl ammonium bromide. These detergents totally dismpt membranes and denature the protein by breaking protein-protein interactions.
  • Non denaturing detergents can be divided into non-anionic detergents such as Triton®X-100, bile salts such as cholates and zwitterionic detergents such as CHAPS. Zwitterionics contain both cationic and anion groups in the same molecule, the positive electric charge is neutralized by the negative charge on the same or adjacent molecule.
  • Diafiltration, or buffer exchange, using ultrafilters is an ideal way for removal and exchange of salts, sugars, non-aqueous solvents separation of free from bound species, removal of material of low molecular weight, or rapid change of ionic and pH environments.
  • Microsolutes are removed most efficiently by adding solvent to the solution being ultrafiltered at a rate equal to the ultrafiltration rate. This washes microspecies from the solution at constant volume, purifying the retained species.
  • the present invention utilizes a diafiltration step to exchange the buffer of the vims supernatant prior to Benzonase ® treatment.
  • Enhancers were originally detected as genetic elements that increased transcription from a promoter located at a distant position on the same molecule of DNA. This ability to act over a large distance had little precedent in classic studies of prokaryotic transcriptional regulation. Subsequent work showed that regions of DNA with enhancer activity are organized much like promoters. That is, they are composed of many individual elements, each of which binds to one or more transcriptional proteins.
  • the liposome may be complexed with a hemagglutinating vims (HVJ). This has been shown to facilitate fusion with the cell membrane and promote cell entry of liposome-encapsulated DNA (Kaneda et al, 1989).
  • the liposome may be complexed or employed in conjunction with nuclear nonhistone chromosomal proteins (HMG-1) (Kato et al, 1991).
  • HMG-1 nuclear nonhistone chromosomal proteins
  • the liposome may be complexed or employed in conjunction with both HVJ and HMG-1. In that such expression constmcts have been successfully employed in transfer and expression of nucleic acid in vitro and in vivo, then they are applicable for the present invention.
  • ion-exchange chromatography The principle of ion-exchange chromatography is that charged molecules adsorb to ion exchangers reversibly so that molecules can be bound or eluted by changing the ionic environment. Separation on ion exchangers is usually accomplished in two stages: first, the substances to be separated are bound to the exchanger, using conditions that give stable and tight binding; then the column is eluted with buffers of different pH, ionic strength, or composition and the components of the buffer compete with the bound material for the binding sites.
  • the Benzonase treated vims solution was purified using IEC. Strong anionic resin Toyopearl SuperQ 650M (Tosohaas) was used for the purification.
  • a FPLC system (Pharmacia) with a XK16 column (Pharmacia) were used for the initial method development. Further scale-up studies were carried out using a BioPilot system (Pharmacia) with a XK 50 column (Pharmacia). Briefly, the resin was packed into the columns and sanitized with 1 N NaOH, then charged with buffer B which was followed by conditioning with buffer A. Buffers A and B were composed of 20 mM

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PCT/US1997/021504 1996-11-20 1997-11-20 An improved method for the production and purification of adenoviral vectors Ceased WO1998022588A2 (en)

Priority Applications (8)

Application Number Priority Date Filing Date Title
CN971812543A CN1244215B (zh) 1996-11-20 1997-11-20 改进的腺病毒载体生产和纯化方法
DE69737107T DE69737107T2 (de) 1996-11-20 1997-11-20 Ein verbessertes verfahren zur produktion und reinigung von adenoviralen vektoren
EP97950677A EP0968284B1 (en) 1996-11-20 1997-11-20 An improved method for the production and purification of adenoviral vectors
AU53617/98A AU732703B2 (en) 1996-11-20 1997-11-20 An improved method for the production and purification of adenoviral vectors
JP52396598A JP4492826B2 (ja) 1996-11-20 1997-11-20 アデノウイルスベクターの産生および精製のための改良された方法
BR9713368-0A BR9713368A (pt) 1996-11-20 1997-11-20 Processo melhorado para a produção e a purificação de vetores adenovirais
CA2272820A CA2272820C (en) 1996-11-20 1997-11-20 An improved method for the production and purification of adenoviral vectors
NO992389A NO992389L (no) 1996-11-20 1999-05-19 Forbedret fremgangsmÕte for fremstilling og rensning av adenovirale vektorer

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US3132996P 1996-11-20 1996-11-20
US60/031,329 1996-11-20

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WO1998022588A2 true WO1998022588A2 (en) 1998-05-28
WO1998022588A3 WO1998022588A3 (en) 1998-10-15
WO1998022588A9 WO1998022588A9 (en) 1998-12-23

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PCT/US1997/021504 Ceased WO1998022588A2 (en) 1996-11-20 1997-11-20 An improved method for the production and purification of adenoviral vectors

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US (7) US6194191B1 (enExample)
EP (3) EP0968284B1 (enExample)
JP (1) JP4492826B2 (enExample)
KR (1) KR100503701B1 (enExample)
CN (1) CN1244215B (enExample)
AT (2) ATE348155T1 (enExample)
AU (1) AU732703B2 (enExample)
BR (1) BR9713368A (enExample)
CA (1) CA2272820C (enExample)
DE (1) DE69737107T2 (enExample)
ES (2) ES2383640T3 (enExample)
NO (1) NO992389L (enExample)
NZ (1) NZ335947A (enExample)
WO (1) WO1998022588A2 (enExample)

Cited By (82)

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WO2000029024A1 (en) * 1998-11-16 2000-05-25 Introgen Therapeutics, Inc. Formulation of adenovirus for gene therapy
WO2000032754A1 (en) * 1998-12-01 2000-06-08 Introgen Therapeutics, Inc. An improved method for the production and purification of adenoviral vectors
FR2788064A1 (fr) * 1998-12-31 2000-07-07 Aventis Pharma Sa Methode de separation de particules virales
WO2000040702A1 (fr) * 1998-12-31 2000-07-13 Aventis Pharma S.A. Methode de separation de particules virales
US6210922B1 (en) * 1998-11-30 2001-04-03 National Research Council Of Canada Serum free production of recombinant proteins and adenoviral vectors
US6261823B1 (en) 1996-12-13 2001-07-17 Schering Corporation Methods for purifying viruses
WO2001077304A1 (en) * 2000-04-07 2001-10-18 Genvec, Inc. Method of producing adenoviral vector stocks
WO2001048155A3 (en) * 1999-12-29 2002-01-03 Genzyme Corp Method using filtration aids for the separation of virus vectors from nucleic acids and other cellular contaminants
US6544769B1 (en) 1996-12-13 2003-04-08 Schering Corporation Compostions comprising viruses and methods for concentrating virus preparations
WO2003093463A1 (en) * 2002-04-30 2003-11-13 Oncolytics Biotech Inc. Improved viral purification methods
EP1371723A1 (en) * 2002-06-12 2003-12-17 Procorde GmbH Process for preparing an adenovirus-containing preparation
US6689600B1 (en) 1998-11-16 2004-02-10 Introgen Therapeutics, Inc. Formulation of adenovirus for gene therapy
US6795585B1 (en) 1999-07-16 2004-09-21 Eastman Kodak Company Representing digital images in a plurality of image processing states
JP2005512565A (ja) * 2001-12-20 2005-05-12 バヴァリアン・ノルディック・アクティーゼルスカブ 感染細胞からのポックスウイルスの採取および精製法
EP1506287A4 (en) * 2002-05-14 2005-06-08 Merck & Co Inc PROCESS FOR CLEANING ADENOVIRUS
WO2005080556A3 (en) * 2004-02-23 2006-02-23 Crucell Holland Bv Virus purification methods
EP1492890A4 (en) * 2002-03-29 2006-10-18 Merck & Co Inc EXPLOSIVE PROCEDURE FOR THE PRODUCTION OF ADENOVIRUS AND ADENOVIRUS STEM MIXTURES
WO2006108707A1 (en) * 2005-04-11 2006-10-19 Crucell Holland B.V. Virus purification using ultrafiltration
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US7264958B1 (en) 1999-02-22 2007-09-04 Transgene, S.A. Method for obtaining a purified viral preparation
US7319002B2 (en) 2001-08-08 2008-01-15 The Trustees Of The University Of Pennsylvania Method for purification of viral vectors having proteins which bind sialic acid
CN100362095C (zh) * 2001-03-27 2008-01-16 沃泰克斯药物股份有限公司 用于hcv感染的组合物和方法
EP1925626A1 (en) 2003-07-21 2008-05-28 Transgene S.A. Novel multifunctional cytokines
US7445930B2 (en) 1996-11-20 2008-11-04 Introgen Therapeutics Inc. Method for the production and purification of adenoviral vectors
WO2009058564A2 (en) 2007-11-01 2009-05-07 Maxygen, Inc. Immunosuppressive polypeptides and nucleic acids
WO2011015656A2 (en) 2009-08-07 2011-02-10 Transgene Sa Composition for treating hbv infection
US7901921B2 (en) 2004-10-22 2011-03-08 Oncolytics Biotech Inc. Viral purification methods
WO2011045381A1 (en) 2009-10-15 2011-04-21 Crucell Holland B.V. Process for adenovirus purification from high cell density cultures
WO2011045378A1 (en) 2009-10-15 2011-04-21 Crucell Holland B.V. Method for the purification of adenovirus particles
WO2011098592A1 (en) 2010-02-15 2011-08-18 Crucell Holland B.V. Method for the production of ad26 adenoviral vectors
EP2390340A2 (en) 2007-01-30 2011-11-30 Transgene SA vector encoding Papillomavirus E1 and E2 polypeptides with reduced percentage of identity
WO2012038367A1 (en) 2010-09-20 2012-03-29 Crucell Holland B.V. Therapeutic vaccination against active tuberculosis
WO2012041669A1 (en) 2010-09-27 2012-04-05 Crucell Holland B.V. Heterologous prime boost vaccination regimen against malaria
WO2012072816A1 (en) 2010-12-03 2012-06-07 Texcell Method for determining the titre of viruses by using infectious standards
WO2012076715A1 (en) 2010-12-09 2012-06-14 Institut Pasteur Mgmt-based method for obtaining high yield of recombinant protein expression
EP2334328A4 (en) * 2008-09-24 2012-08-29 Medimmune Llc METHOD FOR CLEANING VIRUSES
EP2535355A2 (en) 2005-03-23 2012-12-19 Genmab A/S Antibodies against CD38 for treatment of multiple myeloma
WO2013007772A1 (en) 2011-07-12 2013-01-17 Transgene Sa Hbv polymerase mutants
US8357376B2 (en) 2006-09-15 2013-01-22 Memimmune, LLC Method of purifying influenza virus and removing MDCK cell DNA contaminants
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WO2013045668A2 (en) 2011-09-29 2013-04-04 Transgene Sa Immunotherapy composition and regimen for treating hepatitis c virus infection
WO2013074501A1 (en) 2011-11-14 2013-05-23 Crucell Holland B.V. Heterologous prime-boost immunization using measles virus-based vaccines
WO2013135615A1 (en) 2012-03-12 2013-09-19 Crucell Holland B.V. Batches of recombinant adenovirus with altered terminal ends
WO2013139916A1 (en) 2012-03-22 2013-09-26 Crucell Holland B.V. Vaccine against rsv
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