Polymeric Nanoparticles Containing Conjugated Phospholipase A2 for Nonviral Gene Delivery

Polyethylenimine (PEI) was conjugated to phospholipase A2 (PLA2) in an effort to improve transfection efficiency. PLA2 was conjugated to PEI using EDC as a coupling reagent. The activity of enzyme in the conjugate was measured. DNA condensation ability of the conjugate to polymer was determined. The...

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Bibliographic Details
Published in:Molecular pharmaceutics 2010-08, Vol.7 (4), p.1090-1097
Main Authors: Le, Huong T, Rao, Gururaj A, Hirko, Aaron C, Hughes, Jeffrey A
Format: Article
Language:English
Subjects:
Cell Line
Cell Survival
Gene Transfer Techniques
Hep G2 Cells
Humans
Models, Theoretical
Nanoparticles - chemistry
Phospholipases A2 - chemistry
Phospholipases A2 - metabolism
Polyethyleneimine - chemistry
Polymers - chemical synthesis
Polymers - chemistry
Transfection
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Summary:Polyethylenimine (PEI) was conjugated to phospholipase A2 (PLA2) in an effort to improve transfection efficiency. PLA2 was conjugated to PEI using EDC as a coupling reagent. The activity of enzyme in the conjugate was measured. DNA condensation ability of the conjugate to polymer was determined. The resultant nanoparticles were characterized by dynamic and electrophoretic light scattering. Two reporter genes were used to evaluate transfection efficiency in human embryonic kidney (HEK293) and human hepatoma (HepG2) cell lines. Conjugate was shown to retain PLA2 activity and its ability to condense plasmid DNA, resulting in nanoparticles of a similar size to native PEI. The results demonstrated at N/P ratios of 15 and 20 showed 13- and 8-fold increase in transfection efficiency, respectively, compared to the maximum transfection efficiency of PEI (N/P ratio of 5) in the whole range of N/P ratios tested, from 5 to 60 in HepG2 cells. Toxicity studies in HepG2 cells showed uncomplexed conjugate had similar toxicity as PEI, and when complexed with DNA the conjugate had a significantly reduced toxicity. The results clearly indicate the potential for this approach to improve efficiencies of nonviral gene delivery vectors.
ISSN:1543-8384
1543-8392
DOI:10.1021/mp900192p