Large-scale production, purification and crystallization of wild-type adeno-associated virus-2

Qing Xie, Joan Hare, Joy Turnigan, Michael S. Chapman

Research output: Contribution to journalArticle

35 Scopus citations

Abstract

Adeno-associated virus-2 (AAV-2) has long been recognized as a potential vector for human gene therapy. Although much progress has been made in the molecular virology of AAV-2, structural studies of AAV-2 have been hampered by the low efficiency of virus production in culture, the low purity of preparations, and the low solubility of pure virus particles in solution. Methods of larger scale AAV-2 production have been developed through adaptation to suspension culture and re-optimization of the times of infection and transfection with respect to particle production. The methods allow the purification of 10 mg (∼10 15 particles) of AAV-2 per preparation at ∼99% purity as judged by SDS-PAGE. This was sufficient for the screening of conditions for the formation of diffraction-grade crystals, ultimately leading to an atomic structure for AAV-2.

Original languageEnglish (US)
Pages (from-to)17-27
Number of pages11
JournalJournal of Virological Methods
Volume122
Issue number1
DOIs
StatePublished - Dec 1 2004

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Keywords

  • Gene therapy
  • Vector

ASJC Scopus subject areas

  • Virology

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