Mechanistic modeling explains the production dynamics of recombinant adeno-associated virus with the baculovirus expression vector system
Name
1-s2.0-S2329050123000815-main.pdf
Description
Published version
Size
4.28 MB
Format
Adobe PDF
Checksum (MD5)
ca555f5a0a1ae6adf5437a035f72376e
Author(s)
Destro, Francesco
Joseph, John
Srinivasan, Prasanna
Kanter, Joshua M
Neufeld, Caleb
Wolfrum, Jacqueline M
Barone, Paul W
Springs, Stacy L
Sinskey, Anthony J
Cecchini, Sylvain
Date Issued
September 14, 2023
Journal
Molecular Therapy – Methods & Clinical Development
Publisher
Elsevier BV
Citation
Destro, Francesco, Joseph, John, Srinivasan, Prasanna, Kanter, Joshua M, Neufeld, Caleb et al. 2023. "Mechanistic modeling explains the production dynamics of recombinant adeno-associated virus with the baculovirus expression vector system." Molecular Therapy – Methods & Clinical Development, 30.
Version
Final published version
Abstract
Current manufacturing processes for recombinant adeno-associated viruses (rAAVs) have less-than-desired yields and produce significant amounts of empty capsids. The increasing demand and the high cost of goods for rAAV-based gene therapies motivate development of more efficient manufacturing processes. Recently, the US Food and Drug Administration (FDA) approved the first rAAV-based gene therapy product manufactured in the baculovirus expression vector system (BEVS), a technology that demonstrated production of high titers of full capsids. This work presents a first mechanistic model describing the key extracellular and intracellular phenomena occurring during baculovirus infection and rAAV maturation in the BEVS. The model predictions are successfully validated for in-house and literature experimental measurements of the vector genome and of structural and non-structural proteins collected during rAAV manufacturing in the BEVS with the TwoBac and ThreeBac constructs. A model-based analysis of the process is carried out to identify the bottlenecks that limit full capsid formation. Vector genome amplification is found to be the limiting step for rAAV production in Sf9 cells using either the TwoBac or ThreeBac system. In turn, vector genome amplification is hindered by limiting Rep78 levels. Transgene and non-essential baculovirus protein expression in the insect cell during rAAV manufacturing also negatively influences the rAAV production yields.
MIT Department
Massachusetts Institute of Technology. Department of Chemical Engineering
Massachusetts Institute of Technology. Center for Biomedical Innovation
Massachusetts Institute of Technology. Department of Biology
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Creative Commons Attribution-NonCommercial-NoDerivs
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DOI of Published Version
10.1016/j.omtm.2023.05.019
