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Gene therapy has the potential to provide safe and targeted therapies for a variety of diseases. A range of intracellular gene delivery vehicles have been proposed for this purpose. Non-viral vectors are a particularly attractive option and among them cationic peptides have emerged as promising candidates. For the pharmaceutical formulation and application to clinical studies it is necessary to quantify the amount of pDNA condensed with the delivery system. There is a severe deficiency in this area, thus far no methods have been reported specifically for pDNA condensed with cationic peptide to form nanoparticles. The current study seeks to address this and describes the evaluation of a range of disruption agents to extract DNA from nanoparticles formed by condensation with cationic fusogenic peptides RALA and KALA. Only proteinase K exhibited efficient and reproducible results and compatibility with the PicoGreen reagent based quantification assay. Thus we report for the first time a simple and reliable method that can quantify the pDNA content in pDNA cationic peptide nanoparticles.

Original publication

DOI

10.1016/j.jpba.2014.07.036

Type

Journal article

Journal

J Pharm Biomed Anal

Publication Date

11/2014

Volume

100

Pages

236 - 242

Keywords

Cationic peptide, DNA quantification, KALA nanoparticles, PicoGreen, Proteinase K, Cations, DNA, DNA-Binding Proteins, Endopeptidase K, Fluorescent Dyes, Gene Transfer Techniques, Nanoparticles, Nucleic Acid Conformation, Organic Chemicals, Particle Size, Peptides, Protein Conformation, Sodium Dodecyl Sulfate, Temperature, Time Factors