Correlation of Amine Number and pDNA Binding Mechanism for Trehalose-Based Polycations
| Autor: | Theresa M. Reineke, Karina Kizjakina, Lisa E. Prevette, Matthew Lawrence Lynch |
|---|---|
| Rok vydání: | 2008 |
| Předmět: |
Circular dichroism
Stereochemistry Static Electricity Calorimetry Gene delivery chemistry.chemical_compound Spectroscopy Fourier Transform Infrared Polyamines Electrochemistry General Materials Science Amines Particle Size Protein secondary structure Spectroscopy Molecular Structure Chemistry Circular Dichroism Titrimetry Trehalose Water Isothermal titration calorimetry DNA Surfaces and Interfaces Condensed Matter Physics Polyelectrolytes Molecular Weight Solutions Potentiometry Biophysics Thermodynamics Amine gas treating Ethidium bromide Plasmids Macromolecule |
| Zdroj: | Langmuir. 24:8090-8101 |
| ISSN: | 1520-5827 0743-7463 |
| DOI: | 10.1021/la800120q |
| Popis: | Glycopolymers with repeat units comprised of the disaccharide trehalose and an oligoamine of increasing amine have been previously synthesized by our group and shown to efficiently deliver pDNA (plasmid DNA) to HeLa cells while remaining relatively nontoxic. Complexes formed between the most amine-dense of these polycations and pDNA were also found to be relatively stable in serum and have low aggregation, which is desirable for in vivo gene delivery. To lend insight into these interesting results, this study was aimed at investigating the binding strength and mechanism of interaction between these macromolecules, via isothermal titration calorimetry (ITC) and ethidium bromide exclusion assays. The size of these pDNA-polymer complexes, or polyplexes, at various states of formation was determined through light scattering and zeta-potential measurements. Varying degrees of pDNA secondary structure change occurred upon interaction with the polymers, as evidenced by circular dichroism spectra through increasing molar ratios of polymer amine to DNA phosphate, and Fourier transform infrared (FT-IR) results demonstrated stronger electrostatic binding with the phosphate backbone with the least amine-dense of the series. It was concluded that, depending on the number of secondary amines in the repeat unit, these polymers interact with pDNA via different mechanisms with varying extents of electrostatic interaction and hydrogen bonding. These differing mechanisms may affect the ability of trehalose to serve as a deterrent against aggregation in serum conditions and lend insight into the roles of polymer-pDNA binding during the complex transfection process. |
| Databáze: | OpenAIRE |
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