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Research-grade viral vector production

Gene and cell therapy manufacturing services

We can support the early development of your gene therapy and ensure a smooth transition to clinical-grade manufacturing. We offer research/preclinical-grade virus production services for:

  • Retrovirus
  • Lentivirus
  • Adeno-associated virus (AAV)
  • Adenovirus
Retrovirus Lentivirus Adeno-associated virus Adenovirus

Retrovirus  

Our research-grade retrovirus production service lets you choose from multiple expression plasmid vectors depending on the expression type you desire. Stable-cell-line and iPS-cell generation are also available.

Expression plasmid vector options
Expression type
  • High titer (pDON-AI-2)
  • High expression (pMEI-5)
  • Balanced (pDON-5)
Control vector
  • Not available
Virus envelope
  • Amphotropic or ecotropic

Retroviral vector production


  1. Plasmid construction and preparation
    • Cloning of your gene of interest (GOI) into the expression plasmid vector of your choice
    • Large-scale preparation of plasmid DNA for retroviral vector production
  2. Producer cell production (optional)
    • Recommended when performing retroviral vector transduction to human PBMCs.
  3. Retroviral vector production
    • Transfection of 293T cells with plasmid DNA (e.g., pDON-5-GOI and packaging plasmid DNA)
    • Transient expression of retroviral vector and virus collection
    • Viral titration by real-time PCR
  4. Large-scale retroviral vector production
    • Production of retroviral vector at the requested scale
  5. Stable-cell-line generation (optional)
    • Transfection of target cell line using the retroviral vector
    • Antibiotic selection of cells expressing GOI
    • Confirmation of GOI expression by real-time PCR

Lentivirus  

Our research-grade lentivirus production service uses Takara Bio’s proprietary transient lentiviral vector production system. Large-scale preparation and purification by ultracentrifugation are also available.

Expression plasmid vector options
Promoter
  • CMV
  • EF1α
Control vector
  • Not available
Antibiotic resistance
  • Neomycin
  • Hygromycin
  • Puromycin
Virus envelope VSV-G pseudotype

Lentiviral vector production


  1. Plasmid construction and preparation
    • Cloning of your gene of interest (GOI) into expression plasmid DNA (pLVSIN-GOI)
    • Large-scale preparation of plasmid DNA for lentiviral vector production
  2. Lentiviral vector production
    • Transfection of 293T cells with plasmid DNA (pLVSIN-GOI, packaging plasmid DNA)
    • Transient expression of lentiviral vector and virus collection
    • Viral titration by real-time PCR
  3. Large-scale lentiviral vector production and purification
    • Production of lentiviral vector at the requested scale
    • Purification by ultracentrifugation, etc.
  4. Stable-cell-line generation (optional)
    • Transfection of target cell line using the lentiviral vector
    • Antibiotic selection of cells expressing GOI
    • Confirmation of GOI expression by real-time PCR

Adeno-associated virus  

Features of our research-grade AAV production service include:

  • Safer, helper-independent AAV vector with AAVpro Helper Free System
  • Highly purified AAV vector with Takara Bio’s proprietary extraction method
  • Multiple choices of purification method to suit your application
  • Scale-up development for future commercial manufacturing is also available
Expression plasmid vector options
Serotype
  • AAV1
  • AAV2
  • AAV5
  • AAV6
  • Customer-supplied rep-cap plasmid (pRC)
Promoter
  • CMV
Control vector
  • Beta-galactosidase

  • ZsGreen1

AAV vector production


  1. Plasmid construction and preparation
    • Cloning of your gene of interest (GOI) into expression plasmid DNA (pAAV-GOI)
    • Large-scale preparation of plasmid DNA for AAV vector production
  2. AAV vector production (crude)
    • Transfection of three plasmids (pAAV-GOI, pRC, and pHelper) into 293T cells
    • Transient expression of AAV vector
    1. AAV vector purification from cell pellet
      • Collection of AAV vector from cell pellet
      • Purification with AAVpro Purification Kit (All Serotypes)
      • or
    2. AAV vector purification from cell pellet and supernatant
      • Collection of AAV vector from cell pellet and supernatant
      • Purification by affinity chromatography
  3. AAV vector purification with ultracentrifugation (optional)
    • Additional purification with cesium chloride gradient centrifugation

Adenovirus  

Our research-grade adenovirus production service produces high titer, high purity virus for in vivo administration. The adenoviral vector is made from producer cells derived from a single clone.

Expression plasmid vector options
Promoter
  • CMV
  • EF1α
Control vector
  • Axcw2

  • AxCAwt2

  • AxCaiACz

Adenoviral vector production


  1. Cosmid vector construction
    • Cloning of your gene of interest (GOI) into cosmid vector
    • Large-scale preparation of cosmid vector for adenoviral vector production
  2. Clonalized adenoviral vector production
    • Transfection of 293 cells with the adenovirus DNA containing GOI digested from cosmid vector
    • Collection of adenoviral vector from cell supernatant (“full-length transfer method”)
    • Viral titration by hexon detection and RCA contamination testing
    • Viral titration by TCID50 is also available
  3. Adenoviral vector purification (for large-scale [>1 L] only)
    • Purification by cesium chloride gradient centrifugation
    • Viral titration by hexon detection and RCA contamination testing
    • Viral titration by TCID50 is also available

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Takara Bio USA, Inc. provides kits, reagents, instruments, and services that help researchers explore questions about gene discovery, regulation, and function. As a member of the Takara Bio Group, TBUSA is part of a company that holds a leadership position in the global market and is committed to improving the human condition through biotechnology. Our mission is to develop high-quality innovative tools and services to accelerate discovery.

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