Compositional regulation of poly(lysine-g-(lactide-b-ethylene glycol))-DNA complexation and stability.
The search for an effective nonviral gene therapy vector has revealed several significant hurdles, such as transient expression and cytotoxicity, that impede the success of these systems. A terpolymer of poly(lysine-g-(lactide-b-ethylene glycol)) [pK-g-p(LL-b-EG)] has been developed, which is capable of stably packaging DNA with significantly less polycation than unmodified polylysine (pK) systems. A comparison of unmodified pK (DP=14, M(w)=2930) to pK grafted with p(LL-b-EG) (M(w tot)=7500) showed that the minimum amine/phosphate ratio (N:P(min)) needed for complete DNA condensation was reduced by 50%. However, when the molecular weight of pK was reduced (DP=4, M(w)=838), there was evidence of terpolymer interference with DNA condensation. Increasing the number of grafted p(LL-b-EG) chains produced a similar result of incomplete DNA condensation. All terpolymer formulations produced complexes with DNA that had greater resistance to salt-induced dissociation and short-term exchange with excess DNA. Terpolymer-DNA complexes exhibited approximately zero-order plasmid release profiles over a period of 6 weeks. The rate of release was dependent on the complex N:P ratio as well as the molecular weights of pLL and pK. The ability to use terpolymer composition to control complex stability and controlled release can provide a means for system optimization for sustained expression profiles of exogenous DNA.
Park
Susan
S
Departments of Bioengineering and Materials Science and Engineering, University of California at Berkeley, 370 Hearst Memorial Mining Building, Berkeley, CA 94720-1760, USA.
Healy
Kevin E
KE
D004247:DNA / Q000737:chemistry*; D004269:DNA, Bacterial / Q000378:metabolism; D003692:Delayed-Action Preparations / Q000737:chemistry; D004355:Drug Stability; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D008970:Molecular Weight; D010957:Plasmids / Q000378:metabolism; D011092:Polyethylene Glycols / Q000737:chemistry*
D004269:DNA, Bacterial; D003692:Delayed-Action Preparations; C478636:poly(lysine-g-(lactide-b-ethylene glycol)); D011092:Polyethylene Glycols; D004247:DNA