Differential amplification of adenovirus vectors by flanking the packaging signal with attB/attP-ΦC31 sequences: Implications for helper-dependent adenovirus production

Raul Alba, Patrick Hearing, Assumpció Bosch, Miguel Chillon

Research output: Contribution to journalArticleResearchpeer-review

27 Citations (Scopus)

Abstract

Current strategies to amplify helper-dependent adenovirus, based on excision of the packaging signal, do not routinely reduce helper adenovirus contamination below 1%. Here, we have tested if reducing the efficiency of the packaging process of the helper adenovirus could impair its packaging without affecting helper-dependent adenovirus production. Interestingly, insertion of attB/attP-ΦC31 sequences flanking the packaging signal significantly lengthens adenovirus cycle up to 60 h without reducing virus viability or production yield. This delay occurs in the absence of ΦC31 recombinase indicating that other mechanisms different from excision of packaging signal must be involved. In addition, at 36 h post-coinfection helper-dependent adenovirus are efficiently produced, while production levels of helper attB/attP-modified adenovirus are 100-1000 times lower than controls. Therefore, these results suggest that attB/attP-mediated packaging impairment of the adenovirus genome is an attractive strategy to significantly reduce helper adenovirus contamination in helper-dependent adenovirus preparations, which in turn would facilitate scaling-up processes for clinical grade preparations. © 2007 Elsevier Inc. All rights reserved.
Original languageEnglish
Pages (from-to)51-58
JournalVirology
Volume367
DOIs
Publication statusPublished - 10 Oct 2007

Keywords

  • Adenovirus packaging
  • Helper-dependent adenovirus
  • Recombinase FC31

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