title: Impact of molecular weight and degree of conjugation on the thermodynamics of DNA complexation and stability of polyethylenimine-graft-poly(ethylene glycol) copolymers.
abstract: Poly(ethylene glycol) (PEG) is often conjugated to polyethylenimine (PEI) to provide colloidal stability to PEI-DNA polyplexes and shield charge leading to toxicity. Here, a library of nine cationic copolymers was synthesized by grafting three molecular weights (750, 2000, 5000Da) of PEG to linear PEI at three conjugation ratios. Using isothermal titration calorimetry, we have quantified the thermodynamics of the associations between the copolymers and DNA and determined the extent to which binding is hindered as a function of PEG molecular weight and conjugation ratio. Low conjugation ratios of 750Da PEG to PEI resulted in little decrease in DNA affinity, but a significant decrease-up to two orders of magnitude-was found for the other copolymers. We identified limitations in determination of affinity using indirect assays (electrophoretic mobility shift and ethidium bromide exclusion) commonly used in the field. Dynamic light scattering of the DNA complexes at physiological ionic strength showed that PEI modifications that did not reduce DNA affinity also did not confer significant colloidal stability, a finding that was supported by calorimetric data on the aggregation process. These results quantify the DNA interaction thermodynamics of PEGylated polycations for the first time and indicate that there is an optimum PEG chain length and degree of substitution in the design of agents that have desirable properties for effective in vivo gene delivery.
authors.affiliation: Department of Chemistry, University of St. Thomas, 2115 Summit Ave., St. Paul, MN 55105-1079, United States.
authors.forename: Ryan J
authors.initials: RJ
authors.lastname: Smith
authors.affiliation: Department of Chemistry, University of St. Thomas, 2115 Summit Ave., St. Paul, MN 55105-1079, United States.
authors.forename: Rachel W
authors.initials: RW
authors.lastname: Beck
authors.affiliation: Department of Chemistry, University of St. Thomas, 2115 Summit Ave., St. Paul, MN 55105-1079, United States. Electronic address: lisa.prevette@stthomas.edu.
authors.forename: Lisa E
authors.initials: LE
authors.lastname: Prevette
mesh_terms: D002151:Calorimetry; D003102:Colloids / Q000737:chemistry; D004247:DNA / Q000737:chemistry*; D015394:Molecular Structure; D008970:Molecular Weight; D011092:Polyethylene Glycols / Q000138:chemical synthesis / Q000737:chemistry*; D011094:Polyethyleneimine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry; D013816:Thermodynamics
chemical_list: D003102:Colloids; C544016:poly(ethylene glycol)-co-poly(ethyleneimine); D011092:Polyethylene Glycols; D011094:Polyethyleneimine; D004247:DNA
keywords: DNA binding thermodynamics; DNA complexation; Isothermal titration calorimetry; Poly(ethylene glycol) conjugation; Polyethylenimine gene delivery; Polyplex stability