Novel Liposomal Formulation for Targeted Gene Delivery
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Development of a polyethylene glycol (PEG)-stabilized immunoliposome (PSIL) formulation with high DNA content suitable for in vivo intravenous administration and targeted gene delivery.
Materials and Methods
Plasmid DNA was condensed using 40% ethanol and packaged into neutral PSILs targeted to the mouse transferrin receptor using monoclonal antibodies (MAbs; clones RI7 and 8D3) attached to their PEG maleimide moieties. PSILs size was measured by quasi-elastic light scattering. The targeting capacity of the formulation was determined by transfection of mouse neuroblastoma Neuro 2A (N2A) cells with PSIL-DNA complexes conjugated with either RI7 or 8D3 MAbs.
DNA encapsulation and MAb conjugation efficiencies averaged 71 ± 14% and 69 ± 5% (mean ± SD), respectively. No alteration in mean particle size (< 100 nm) or DNA leakage were found after 48 h storage in a physiological buffer, and the in vivo terminal half-life reached 23.9 h, indicating that the PSIL-DNA formulation was stable. Addition of free RI7 MAbs prevented transfection of N2A cells with PSIL-DNA complexes conjugated with either RI7 or 8D3 MAbs, confirming that the transfection was transferrin receptor-dependent.
The present data suggest that our new PSIL formulation combines molecular features required for targeted gene therapy including high DNA encapsulation efficiencies and vector-specific transient transfection capacity.
Key wordsDNA encapsulation gene therapy liposomes monoclonal antibodies transferrin receptors
bovine serum albumine
disintegrations per minute
herpes simplex virus-1
liquid scintillation counting
molecular weight cut-off
neuroblastoma neuro 2A
phosphate buffered saline
polymerase chain reaction
2,000 da polyethylene glycol
relative light units
quasi-elastic light scattering
standard error mean
severe combined immunodeficiency
simian virus 40
virus production serum-free medium