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Methods for generating high titer helper-free preparations of released recombinant AAV vectors

  • US 6,566,118 B1
  • Filed: 03/15/2000
  • Issued: 05/20/2003
  • Est. Priority Date: 09/05/1997
  • Status: Expired due to Term
First Claim
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1. A method of generating a population of recombinant adeno-associated virus (rAAV) particles, comprising the following steps:

  • a) incubating producer cells in a cell culture medium under conditions comprising a condition that promotes release of rAAV particles into the medium without lysing the cells, wherein the producer cells comprise;

    (i) one or more packaging genes, wherein each said AAV packaging gene encodes an AAV replication or encapsidation protein;

    (ii) a recombinant AAV (rAAV) vector that comprises a heterologous non-AAV polynucleotide flanked by at least one AAV inverted terminal repeat (ITR); and

    (iii) helper virus function for AAV, whereby rAAV virus particles are released from the producer cells into the cell culture medium, and b) harvesting the rAAV Particles from the cell culture medium without lysing the cells.

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