Chemically Defined, High-Density Insect Cell-Based Expression System for Scalable AAV Vector Production
| العنوان: | Chemically Defined, High-Density Insect Cell-Based Expression System for Scalable AAV Vector Production |
|---|---|
| المؤلفون: | Andrey Tovchigrechko, Yasuhiro Ikeda, Claire Dobson, Andrew Park, Susan Wilson, Rebecca A. Halpin, Changshou Gao, Carrie R. Sowers, Albert E. Schmelzer, James H. Kurasawa |
| المصدر: | Molecular Therapy. Methods & Clinical Development Molecular Therapy: Methods & Clinical Development, Vol 19, Iss, Pp 330-340 (2020) |
| بيانات النشر: | Elsevier BV, 2020. |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | 0301 basic medicine, lcsh:QH426-470, viruses, Genetic enhancement, Transgene, Sf9, Biology, Virus, Viral vector, law.invention, 03 medical and health sciences, Transduction (genetics), baculovirus, 0302 clinical medicine, law, Genetics, lcsh:QH573-671, Molecular Biology, insect cell, lcsh:Cytology, AAV, Cell biology, lcsh:Genetics, Chemically defined medium, 030104 developmental biology, 030220 oncology & carcinogenesis, Recombinant DNA, Molecular Medicine, Original Article |
| الوصف: | The recombinant adeno-associated virus (AAV) vector is one of the most utilized viral vectors in gene therapy due to its robust, long-term in vivo transgene expression and low toxicity. One major hurdle for clinical AAV applications is large-scale manufacturing. In this regard, the baculovirus-based AAV production system is highly attractive due to its scalability and predictable biosafety. Here, we describe a simple method to improve the baculovirus-based AAV production using the ExpiSf Baculovirus Expression System with a chemically defined medium for suspension culture of high-density ExpiSf9 cells. Baculovirus-infected ExpiSf9 cells produced up to 5 × 1011 genome copies of highly purified AAV vectors per 1 mL of suspension culture, which is up to a 19-fold higher yield than the titers we obtained from the conventional Sf9 cell-based system. When mice were administered the same dose of AAV vectors, we saw comparable transduction efficiency and biodistributions between the vectors made in ExpiSf9 and Sf9 cells. Thus, the ExpiSf Baculovirus Expression System would support facile and scalable AAV manufacturing amenable for preclinical and clinical applications. Graphical Abstract Large-scale manufacturing is a major challenge for preclinical and clinical AAV vector applications. Ikeda and colleagues describe a simple method using a high-density insect cell-based system to improve AAV vector yield up to 19-fold higher than the conventional Sf9 cell-based system, which would facilitate scalable AAV vector manufacturing. |
| تدمد: | 2329-0501 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::39bcde561465c042ac7f700ec6386eef https://doi.org/10.1016/j.omtm.2020.09.018 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....39bcde561465c042ac7f700ec6386eef |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 23290501 |
|---|