Production of first generation adenovirus vectors: a review

Abstract

In the past decade, adenovirus vectors have generated tremendous interest, especially in gene therapy applications. In the so-called 'first generation' adenovirus vectors, the transgenes are inserted in place of the E1 region, or less often the E3 region. Although second-generation and helperdependent adenovirus vectors will probably prevail in the future in applications that require long-term gene expression, first generation adenovirus vectors will remain very useful in

Chromatographic purification of recombinant adenoviral and adeno-associated viral vectors: methods and implications

“…Modern manufacturing methods allow for production of pure and concentrated Ad stocks required for human gene therapy. [4][5][6] AAV is a small, nonenveloped, icosahedral virus 20-26 nm in diameter and with a linear single-stranded DNA genome of 4.7À6 kb. 7,8 Recombinant adeno-associated viral (rAAV) vectors as a class appear to be the most suitable for applications where persistent transgene expression is essential for achieving a therapeutic goal.…”

“…Construction of rAd genome can be achieved via homologous or Cre-mediated recombination in mammalian cells, or via reconstruction of fulllength viral genome in bacteria (reviewed by Danthinne and Imperiale). 6 Lack of the viral E1 gene product renders rAd replication-deficient. Therefore, amplification of rAd particles is only possible in stable producer cell lines containing an integrated E1 transgene.…”

Viral vectors for in vivo gene transfer in Parkinson's disease: Properties and clinical grade production

“…First generation recombinant Ad vectors have a deletion of the E1A early gene that is responsible for viral growth and induction of late genes. For production of the recombinant form of Ad, the E1 gene product can be provided in trans in another separate plasmid (Danthinne and Imperiale, 2000). This allows for the deletion of the E1A gene which renders the virus replication deficient and allows introduction of therapeutic genes in place of the E1A gene (see Fig.…”

Gutless adenovirus: last-generation adenovirus for gene therapy

“…In order to produce safe and nonreplicative vectors, first-generation adenoviral vectors containing the whole viral genome with the exception of the E1 region were developed. 2 To propagate first-generation adenoviruses, several E1-expressing cell lines have been generated: 293, 3 911, 4 N52.E6 5 and PER.C6. 6 Although E1-deleted vectors cannot replicate in vivo, residual expression from adenoviral genes triggers a cytotoxic T lymphocyte (CTL) immune response towards infected cells, 7 which finally leads to the elimination of transduced cells and, therefore, to the lost of therapeutic gene expression.…”