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Intensified Production of Vaccinia-Based Oncolytics in the High Density Cell Respirator (HDCR) Bioreactor Improves Vaccine Logistics and Economics

Cook, Colin A. and Kang, Seonah and Lu, Jianming and Tai, Yu-Chong and Chatterjee, Saswati and Fong, Yuman (2021) Intensified Production of Vaccinia-Based Oncolytics in the High Density Cell Respirator (HDCR) Bioreactor Improves Vaccine Logistics and Economics. Molecular Therapy, 29 (4, S1). pp. 397-398. ISSN 1525-0016. https://resolver.caltech.edu/CaltechAUTHORS:20210528-141159173

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Abstract

Intensification of cell-based production processes is key to improving oncolytic vaccine logistics and economics by saving on GMP space, time, labor, and feedstock. Many promising vectors, including those based on vaccinia virus (e.g. CF33, JX-594/Pexa-Vec) are still produced using flask-based culture due to the cost, effort, and uncertainty involved in adapting to stirred-tank or perfusion processes. Based on prior success culturing adherent cells (e.g. HEK293, A549, CV-1) to high densities (107-8 cells/mL) using the scalable high-density cell respirator (HDCR) bioreactor, we hypothesized that the platform could support orders-of-magnitude-intensified production of replication competent viruses. Here we report on CF33 virus production as a proof-of-concept for oncolytic virotherapy. Bioreactor cartridges were produced based on a 4-stack of 600 cm² HDCR membranes, featuring a proprietary gas perfusable and permeable microarchitecture that optimally oxygenates cells even at high densities. Media perfusion was feedback-controlled based on glucose measurements in the waste stream. A549 cells were seeded into the bioreactor on microcarriers, expanded 10-fold to mid-10⁷ cells/mL densities, infected with CF33 virus (MOI of 0.1), and harvested 48 hours post-infection. CF33 virus was gradient purified and titered for functional virus by plaque forming assay. Multiple production runs using different strains of CF33 virus validated the reproducibility of the process, as summarized in Table 1. Importantly, cell specific titers (PFU/cell) remained comparable to conventional flask-based production, leading to significant intensification due to the higher cell densities supported in the HDCR bioreactor. Volumetric productivity is on the order of 100× that of cell factories. The efficient usage of media due to the gas-media decoupled operation of the HDCR bioreactor enables gradient-purified virus costs of around $500/1010 PFU. This is significantly below the $10,000/1010 PFU charged by CMOs and supports our mission of democratizing access to life saving medicines. The straightforward adaptation of CF33 virus production from a flask-based to a high yield, intensified process highlights the logistical and economical advantage of the HDCR platform for oncolytics.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1016/j.ymthe.2021.04.019DOIAbstracts
ORCID:
AuthorORCID
Cook, Colin A.0000-0002-6283-5105
Tai, Yu-Chong0000-0001-8529-106X
Additional Information:© 2021 American Society of Gene & Cell Therapy. Available online 27 April 2021.
Issue or Number:4, S1
Record Number:CaltechAUTHORS:20210528-141159173
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20210528-141159173
Official Citation:ASGCT Annual Meeting Abstracts, Molecular Therapy, Volume 29, Issue 4, Supplement 1, 2021, Pages 1-427, ISSN 1525-0016, https://doi.org/10.1016/j.ymthe.2021.04.019. (https://www.sciencedirect.com/science/article/pii/S1525001621002069)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:109307
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:28 May 2021 22:13
Last Modified:28 May 2021 22:13

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