FDA Perspectives on the Use of the Adenovirus Reference Material

Simek, Stephanie L.; Byrnes, Andrew P.; Bauer, Steven R.

doi:10.12665/j13.simek

Cited by 25 publications

(10 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…For clinical applications, the FDA Advisory Committee recommends a ratio equal or below 30:1 . In this work the VG: IP ratio obtained was of 5.2 (± 1.9): 1, very favorably meeting the authorities requirements . TEM analysis confirmed the presence, integrity and expected morphology of the produced virus.…”

Section: Resultssupporting

confidence: 59%

Establishing Suspension Cell Cultures for Improved Manufacturing of Oncolytic Adenovirus

Moreira

Silva

Sousa

et al. 2020

Biotechnology Journal

View full text Add to dashboard Cite

Recent clinical trials have shown the potential of oncolytic adenoviruses as a cancer immunotherapy. A successful transition of oncolytic adenovirus to clinical applications requires efficient and good manufacturing practice compatible production and purification bioprocesses. Suspension cultures are preferable for virus production as they can reduce process costs and increase product quality and consistency. This work describes the adaptation of the A549 cell line to suspension culture in serum‐reduced medium validated by oncolytic adenovirus production in stirred tank bioreactor. Cell concentrations up to 3 × 106 cells mL−1 are obtained during the production process. At harvest 1.4 × 1010 infectious particles mL−1 and 6.9 ± 1.1 × 1010 viral genome mL−1 are obtained corresponding to a viral genome: infectious particles ratio of 5.2 (± 1.9): 1 confirming the virus quality. Overall, the suspension characteristics of these A549 cells support an easily scalable, less time‐consuming, and more cost‐effective process for expanded success in the use of oncolytic viruses for cancer therapy.

show abstract

Section: Resultssupporting

confidence: 59%

Establishing Suspension Cell Cultures for Improved Manufacturing of Oncolytic Adenovirus

Moreira

Silva

Sousa

et al. 2020

Biotechnology Journal

View full text Add to dashboard Cite

show abstract

“…46,47 A key parameter in processes related with the production of vectors for gene therapy is the ratio of viral genomes per infectious particles (vg/ip). For this type of application, an FDA Advisory Committee recommended a targeted vg/ip ratio below 30 (Simek et al, 2002). 48 In this study, the ratios vg/ip estimated for the ST and VW-PBS bioreactors were 151 6 37 and 64 6 30 vg/ip, respectively ( Figure 3B).…”

Section: A549 Cell Growth and Onco-ad5 Productionmentioning

confidence: 70%

“…For this type of application, an FDA Advisory Committee recommended a targeted vg/ip ratio below 30 (Simek et al, 2002). 48 In this study, the ratios vg/ip estimated for the ST and VW-PBS bioreactors were 151 6 37 and 64 6 30 vg/ip, respectively ( Figure 3B). Although these values are well above the recommended target, it is important to highlight that the PBS-VW bioreactor induced a vg/ip ratio 2.5 times lower than the ST bioreactor.…”

Section: A549 Cell Growth and Onco-ad5 Productionmentioning

confidence: 70%

Production of oncolytic adenovirus and human mesenchymal stem cells in a single‐use, Vertical‐Wheel bioreactor system: Impact of bioreactor design on performance of microcarrier‐based cell culture processes

Sousa

Silva

Giroux

et al. 2015

Biotechnology Progress

View full text Add to dashboard Cite

Anchorage-dependent cell cultures are used for the production of viruses, viral vectors, and vaccines, as well as for various cell therapies and tissue engineering applications. Most of these applications currently rely on planar technologies for the generation of biological products. However, as new cell therapy product candidates move from clinical trials towards potential commercialization, planar platforms have proven to be inadequate to meet large-scale manufacturing demand. Therefore, a new scalable platform for culturing anchorage-dependent cells at high cell volumetric concentrations is urgently needed. One promising solution is to grow cells on microcarriers suspended in single-use bioreactors. Toward this goal, a novel bioreactor system utilizing an innovative Vertical-Wheel™ technology was evaluated for its potential to support scalable cell culture process development. Two anchorage-dependent human cell types were used: human lung carcinoma cells (A549 cell line) and human bone marrow-derived mesenchymal stem cells (hMSC). Key hydrodynamic parameters such as power input, mixing time, Kolmogorov length scale, and shear stress were estimated. The performance of Vertical-Wheel bioreactors (PBS-VW) was then evaluated for A549 cell growth and oncolytic adenovirus type 5 production as well as for hMSC expansion. Regarding the first cell model, higher cell growth and number of infectious viruses per cell were achieved when compared with stirred tank (ST) bioreactors. For the hMSC model, although higher percentages of proliferative cells could be reached in the PBS-VW compared with ST bioreactors, no significant differences in the cell volumetric concentration and expansion factor were observed. Noteworthy, the hMSC population generated in the PBS-VW showed a significantly lower percentage of apoptotic cells as well as reduced levels of HLA-DR positive cells. Overall, these results showed that process transfer from ST bioreactor to PBS-VW, and scale-up was successfully carried out for two different microcarrier-based cell cultures. Ultimately, the data herein generated demonstrate the potential of Vertical-Wheel bioreactors as a new scalable biomanufacturing platform for microcarrier-based cell cultures of complex biopharmaceuticals.

show abstract

“…26 Additionally, following transduction ex vivo, the transduced dermal MOs were extensively washed and maintained in organ culture for 9 days to remove most capsid proteins before implantation. Because the reduction of HuEPO secretion from the transduced dermal tissue kept in organ culture was slower than the decline of circulating HuEPO in the implanted mice, we hypothesized that implantation in the SCID mice was somewhat detrimental to the implanted tissue .…”

Section: Human Gene Therapy Clinical Developmentmentioning

confidence: 99%

Preclinical and preliminary clinical evaluation of genetically transduced dermal tissue implants for the sustained secretion of erythropoietin and interferon α

Shapir¹,

Miari²,

Blum³

et al. 2015

Human Gene Therapy Clinical Development

View full text Add to dashboard Cite

Protein drugs are currently delivered by bolus injection and although treatment frequently is successful, these methods also have major drawbacks, which call for the development of alternative technologies allowing prolonged delivery of these drugs. We developed a new ex vivo gene therapy platform called Transduced Autologous Restorative Gene Therapy (TARGT) for sustained long term production and secretion of autologous therapeutic proteins. A biopsy of dermal tissue taken from the patient is transduced ex vivo with a viral vector encoding the required gene under a constitutive promoter. Following measurement of protein secretion ex vivo, the transduced dermal tissue is implanted back into the patient, where it secretes the therapeutic protein into the circulation for several months or longer. A major hurdle to this approach is potential immunogenicity of the transduced tissue following implantation. In this paper we describe the preclinical and early clinical development of this technology, which allowed overcoming these hurdles. To that end, we have used the helper dependent (HD) adenoviral vector with newly designed expression cassette containing genetic elements to optimize transgene expression. Moreover, we have developed procedures for TARGT tissue implantation, with measures to improve engraftment and reduce inflammation and rejection. Implantation of human TARGT to immune-deficient SCID mice indicated long-term production of active proteins in the blood. Preliminary results of a clinical trial from two anemic end-stage renal disease patients, implanted with TARGTs expressing the human erythropoietin (EPO) gene, demonstrated prolonged secretion with physiologic blood level of the hormone and Hemoglobin maintenance in the desired range, for a period of at least five months without exogenous EPO administration. We believe that the TARGT technology has the potential to become a platform for the sustained delivery of therapeutic proteins in various clinical indications.

show abstract

FDA Perspectives on the Use of the Adenovirus Reference Material

Cited by 25 publications

References 0 publications

Establishing Suspension Cell Cultures for Improved Manufacturing of Oncolytic Adenovirus

Establishing Suspension Cell Cultures for Improved Manufacturing of Oncolytic Adenovirus

Production of oncolytic adenovirus and human mesenchymal stem cells in a single‐use, Vertical‐Wheel bioreactor system: Impact of bioreactor design on performance of microcarrier‐based cell culture processes

Preclinical and preliminary clinical evaluation of genetically transduced dermal tissue implants for the sustained secretion of erythropoietin and interferon α

Contact Info

Product

Resources

About