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- Vector Manufacturing for Gene & Cell Therapy
A previous project of “new manufacturing technologies on large scale production of viral vectors for gene and cell therapy” was completed in FY2023. Now, a next project on the research and development entitled “Integrated Development of Viral Vector Manufacturing Technology” in the AMED program as “Fundamental Technology Development Project for Industrialization of Regenerative Medicine and Gene Therapy (Research Project for Accelerating Development of Gene Therapy)” has been started from FY2024. In order to accelerate the development of gene and cell therapy drugs, industry, government, and academia that possess related technologies in Japan will come together to promote the project aiming to conduct clinical trials（ including investigator-initiated clinical trials）.
Integrated Development of Viral Vector Manufacturing Technology (FY2024-FY2029)
- ① Development of original cell lines
- Characterization of HAT-cell (Human Amniotic epithelial cell line for gene & cell Therapy) developed by MAB and development of HAT-cells as a host cell for viral vector manufacturing
- ② Development of manufacturing process
- Development and optimization of manufacturing process for viral vectors using HAT-cells
- ③ Promotion and acceleration to clinical application
- Development of manufacturing process for therapeutic viral vector for genetic disease
Production process of Viral vectors
(October 1,2024)
Kusatsu Consolidated Lab.
Kawasaki Consolidated Lab.
Osaka Consolidated Lab.
R&D of new technologies on large scale production of viral vectors for gene and therapy (FY2018～2023)
1. R&D subjects
Development of elemental technology
- ① Construction of production cell line suitable for viral vector production
- ② Development of upstream technology
- ③ Development of downstream technology
- ④ Development of analytical technology
- ⑤ Construction of manufacturing technology platform
Development of manufacturing technology platform
2. R&D Organization
(April 1,2024)
3. R&D Results
(1) Establishment and development of a novel cell line, HAT*, derived from human placental tissues**, with rapid cell proliferation and high-yield AAV productivity
The Cell Line Development team of MAB, based in Kawasaki Consolidated Laboratory/Chitose Laboratory Corp. and National Research Institute for Child Health and Development, has successfully established an original HAT cell line as a novel platform for therapeutic viral vector production. To meet the biomanufacturing requirement, HAT cells were adapted to grow in serum-free suspension culture conditions, and clonal HAT cells with full history documentation were isolated. HAT cells exhibit greater or comparable cell proliferation ability and AAV productivity to the commonly used commercially available HEK293 cells, showing high potential applicability for viral vector manufacturing for gene therapy.
HAT cells are available for research or commercial use under certain conditions. Please email ” contact@chitose-bio.com ” for further information.
- *
- HAT: Human Amniotic epithelial cell line for gene and cell Therapy
- **
- Human placental tissues were donated voluntarily from healthy mothers under appropriate informed consent after scheduled cesarean deliveries performed at National Center for Child Health and Development.
[ Proliferation and AAV production capability of the HAT A2C1 cell lines ]
- *
- HAT A2C1： Clonal strain of HAT cells
VPCs 2.0 : Derivative strain of HEK293 cells
HAT A2C1 showed higher proliferation and productivity than HEK293.
AAV vectors were produced by triple transfection under serum-free suspension culture conditions in 125-mL flasks. Genomic titers of AAV2 and AAV5 were quantified by qPCR.
(2) Development of various elemental technologies and creation of an integrated platform for production of viral vectors
By each research institutes participating in this project, elemental technologies related to the production process of viral vectors were examined, and new technologies and products were developed in each process of production cell establish, seed & expansion culture, production culture followed by plasmid transfection, clarification of cell solubilizing solution, affinity, polishing, and other purification processes.
Elemental technologies developed in this project include a culture process using a domestically produced culture reactor (ZACROS), adsorbent (KANEKA) and filtration filters (TORAY) in the clarification process of cell solubilization solution, ion exchange chromatography resin and purification process for separation of full and empty AAV particles (JNC, YMC), and zonal ultracentrifugation purification technology (University of Tokyo).
The series of elemental technologies developed were integrated at the Kusatsu Consolidated Lab., a manufacturing technology development center, to construct a manufacturing process (integrated platform) for AAV viral vectors from 50L-scale culture medium to the purification of full particles. As a result of the full particles measurement, including analysis of genome titer, full/empty particle ratio, capsid size, etc., it was demonstrated that high-quality AAV can be produced at a high yield.
(3) Development of advanced analytical technology and platform for production of viral vectors
We have established the Osaka Consolidated Lab., Osaka University and Kobe University as a center for analytical technology development by equipping them with the instruments and equipment necessary for advanced quality analysis of viral vectors. We also have built a one-stop system and platform for almost all analyses required by the guidelines in Japan, the U.S., and Europe.
The advanced quality analysis technology developed in this project has made it possible to analyze the quality of a smaller quantity of samples than before, enabling analysis and evaluation at each step in the development of the manufacturing process for viral vectors and efficient manufacturing process development on a small scale. It is expected that this analytical technology will facilitate the development of low-cost, rapid, and highly efficient manufacturing processes in the future.
[ Established guality analysis methods ]
