Research-grade viral vector production

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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

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
Producer cell production (optional)
- Recommended when performing retroviral vector transduction to human PBMCs.
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
Large-scale retroviral vector production
- Production of retroviral vector at the requested scale
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

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
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
Large-scale lentiviral vector production and purification
- Production of lentiviral vector at the requested scale
- Purification by ultracentrifugation, etc.
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

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
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
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

Cosmid vector construction
- Cloning of your gene of interest (GOI) into cosmid vector
- Large-scale preparation of cosmid vector for adenoviral vector production
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
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.
United States/Canada: +1.800.662.2566 • Asia Pacific: +1.650.919.7300 • Europe: +33.(0)1.3904.6880 • Japan: +81.(0)77.565.6999
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