Spectroscopic methods for the physical characterization and formulation of nonviral gene delivery systems.
Currently, with the exception of naked DNA formulations, most pharmaceutical preparations of plasmid DNA employ some type of polycationic delivery vector such as synthetic cationic polymers and lipids to enhance delivery. A number of biophysical techniques are readily available for the structural characterization of plasmid DNA within synthetic gene delivery complexes. Here we present applications of ultraviolet (UV) absorption, circular dichroism (CD), infrared (IR), and fluorescence spectroscopies as well as dynamic light scattering to the structural analysis of the oligonucleotide component of nonviral gene delivery vectors. We also illustrate this approach for the investigation of the formulation of lipoplex and polyplex-based gene delivery systems. To summarize such data, we show how the macromolecular complexes can be represented as vectors in a highly dimensional space in which the components of the vector consist of normalized values of experimental parameters measured as a function of different solution conditions such as pH and ionic strength.
Ausar
Salvador F
SF
Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, KS, USA.
Joshi
Sangeeta B
SB
Middaugh
C Russell
CR
D002942:Circular Dichroism / Q000379:methods*; D004247:DNA / Q000008:administration & dosage / Q000737:chemistry; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006863:Hydrogen-Ion Concentration; D008081:Liposomes / Q000737:chemistry*; D046911:Macromolecular Substances; D009994:Osmolar Concentration; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013055:Spectrophotometry, Infrared / Q000379:methods*; D013056:Spectrophotometry, Ultraviolet / Q000379:methods*
D008081:Liposomes; D046911:Macromolecular Substances; D004247:DNA