Development of a method to quantify the DNA content in DNA- cationic peptide nanoparticles

Arvind Jain, Helmy Yusuf, Aditya Pattani, Helen McCarthy, Denise McDonald, Victoria Kett

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

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 languageEnglish
Pages (from-to)236-242
JournalJournal of Pharmaceutical and Biomedical Analysis
Volume100
Early online date09 Aug 2014
DOIs
Publication statusPublished - Nov 2014

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