Display Settings:

Format

Send to:

Choose Destination
    Hum Gene Ther. 2001 Jan 1;12(1):71-6.

    Isolation of highly infectious and pure adeno-associated virus type 2 vectors with a single-step gravity-flow column.

    Source

    Wistar Institute, Philadelphia, PA 19104, USA.

    Abstract

    One of the most promising gene transfer vectors in human clinical trials is AAV2. The quality of the vector preparations is a key element in obtaining reliable and reproducible data in preclinical studies. However, established protocols either result in impure, low infectious virus (CsCl2 gradient centrifugation) or demand a high level of manual and technical skills (CsCl2 gradient centrifugation, iodixanol/heparin or HPLC purification). In this study, we present an easy-to-do single-step column purification (SSCP) of AAV2 by gravity flow based on its affinity to heparin, without ultracentrifugation. Various vector preparations generated by our method reproducibly showed high titers, infectivity, and purity. In vivo, our single-step column-purified AAV2 vectors mediate significantly higher transduction efficiency compared with conventional protocols. Investigators still unsatisfied with previously published techniques or new to the field of AAV production may find in our method an interesting alternative.

    PMID:
    11177544
    [PubMed - indexed for MEDLINE]

      Supplemental Content

      Icon for Mary Ann Liebert, Inc.

      Save items

      loading

      Recent activity

      Your browsing activity is empty.

      Activity recording is turned off.

      Turn recording back on

      See more...
      Write to the Help Desk