Preclinical Proof-of-Concept, Analytical Development, and Commercial Scale Production of Lentiviral Vector in Adherent Cells
Autor: | Heidi Hynynen, Haritha Samaranayake, Ann-Marie Määttä, Jere Kurkipuro, Hanna Leinonen, Anniina J. Valkama, Nigel Parker, Venla Olsson, Seppo Ylä-Herttuala, Hanna P. Lesch, Vesa Turkki, Eevi M. Lipponen, Igor Oruetxebarria |
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Jazyk: | angličtina |
Rok vydání: | 2019 |
Předmět: |
0301 basic medicine
Ganciclovir lcsh:QH426-470 Pharmacology medicine.disease_cause Article Viral vector 03 medical and health sciences bioreactor 0302 clinical medicine lentivirus Glioma glioma Genetics medicine Potency Viability assay lcsh:QH573-671 Molecular Biology business.industry lcsh:Cytology Transfection medicine.disease lcsh:Genetics 030104 developmental biology Herpes simplex virus transfection Thymidine kinase 030220 oncology & carcinogenesis Molecular Medicine production business scale up medicine.drug |
Zdroj: | Molecular Therapy: Methods & Clinical Development, Vol 15, Iss, Pp 63-71 (2019) Molecular Therapy. Methods & Clinical Development |
ISSN: | 2329-0501 |
Popis: | The therapeutic efficacy of a lentiviral vector (LV) expressing the herpes simplex virus thymidine kinase (HSV-TK) was studied in an immunocompetent rat glioblastoma model. Intraperitoneal ganciclovir injections (50 mg/kg/day) were administered for 14 consecutive days, resulting in reduced tumor volumes as monitored by MRI. Survival analyses revealed a significant improvement among the LV-expressing HSV-TK (LV-TK)/ganciclovir-treated animals when compared to non-treated control rats. However, a limiting factor in the use of LV has been the suboptimal small-scale production in flasks. Our aim during the translation phase, prior to entering the final pre-clinical and early clinical phases, was to develop a scalable, robust, and disposable manufacturing process for LV-TKs. We also aimed to minimize future process changes and enable production upscaling to make the process suitable for larger patient populations. The upstream process relies on fixed-bed iCELLis technology and transient plasmid transfection. This is the first time iCELLis 500 commercial-scale bioreactor was used for LV production. A testing strategy to determine the pharmacological activity of LV-TK drug product by measuring cell viability was developed, and the specificity of the potency assay was also proven. In this paper we focus on upstream process development while showing analytical development and the proof-of-concept of LV-TK functionality. Keywords: lentivirus, bioreactor, transfection, production, scale up, glioma |
Databáze: | OpenAIRE |
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