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. 2014 Mar 10;2(1):110–131. doi: 10.3390/biomedicines2010110

Figure 2.

Figure 2

Overview of the production of Helper-dependent Ad vectors (HDAds). (A) A flow chart of the large-scale production of HDAd. The HDAd plasmid DNA (pHDAd) is linearized with the restriction enzyme PmeI before transfection to producer cell, 116 cell overexpressing Cre. HDAds are amplified by serial co-infection of helper virus and subjected to a 10-chamber cell factory. HDAd virions are purified from cell lysate by CsCl ultracentrifugation; (B) Generation of chimeric or different serotype HDAds. HDAd5 encoding transgene expression cassette(s) is co-infected with appropriate helper virus (chimeric or different serotype) in a 10-chamber cell factory, and produced HDAd capsid harbors helper virus dependent capsid.