Methods for rapid PEG-modification of viral vectors, compositions for enhanced gene transduction, compositions with enhanced physical stability, and uses therefor
AssigneeThe Trustees of the University of Pennsylvania
InventorsMaria Croyle, James M. Wilson
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Abstract
A rapid method for modifying a viral capsid or envelope protein with a polyethylene glycol (PEG) is described. Also provided are methods of delivering a molecule using the PEG-modified viruses of the invention. Compositions containing the PEG-modified viruses of the invention, are characterized by improved gene expression, reduced neutralizing antibody and CTL production. Also provided are viral compositions having enhanced physical stability, in which the viruses are lyophilized in a formulation having a 1:1 ratio of sucrose and mannitol are provided.
Claims34
A method for conjugating a recombinant virus with polyethylene glycol to enhance transduction efficiency thereof, said method comprising the steps of:
- a) reacting activated polyethylene glycol and a recombinant virus at ambient temperature for about 1 to about 2 hours at a ratio of about 10:1 polyethylene glycol to virus;
- and
- b) stopping the reaction, thereby obtaining polyethylene glycol conjugated virus.
A method for conjugating a recombinant adeno-associated virus with polyethylene glycol to enhance transduction efficiency thereof, said method comprising the steps of:
- a) reacting activated polyethylene glycol and a recombinant adeno-associated virus at ambient temperature for about 1 to about 2 hours;
- and
- b) stopping the reaction, thereby obtaining polyethylene glycol conjugated adeno-associated virus.
A method for increasing transduction efficiency of a recombinant virus, said method comprising the steps of:
- (a) conjugating a recombinant virus with polyethylene-glycol by
- (i) reacting activated polyethylene glycol and a recombinant virus at ambient temperature for about 1 to about 2 hours at a ratio of about 10:1 polyethylene glycol to virus;
- and
- (ii) stopping the reaction, thereby obtaining polyethylene glycol conjugated virus with increased transduction efficiency;
- and
- (b) delivering the polyethylene-glycol conjugated virus to host cells.
A composition that enhances the physical stability of viral vectors, said composition comprising:
- (a) a recombinant viral vector comprising a molecule for delivery to host cells;
- (b) sucrose;
- and
- (c) mannitol, wherein the ratio of sucrose to mannitol is about 1 to about 1.
A method for increasing transduction efficiency of a recombinant adeno-associated virus, said method comprising the steps of:
- (a) conjugating a recombinant adeno-associated virus with polyethylene-glycol by
- i) reacting activated polyethylene glycol and a recombinant adeno-associated virus at ambient temperature for about 1 to about 2 hours;
- and
- ii) stopping the reaction, thereby obtaining polyethylene glycol conjugated adeno-associated virus;
- and
- (b) delivering the polyethylene-glycol conjugated virus to host cells.
Dependencies are inferred from the claim text.