Rapid and sensitive ethidium bromide fluorescence quenching assay of polyamine conjugate-DNA interactions for the analysis of lipoplex formation in gene therapy

A J Geall, Ian S Blagbrough

Research output: Contribution to journalArticle

125 Citations (Scopus)

Abstract

A rapid and sensitive fluorescent assay method is reported for assessing polyamine conjugate-calf thymus DNA binding affinity using cholesterol polyamine carbamates with ethidium bromide as a probe. A reproducible method has been developed with an optimal excitation wavelength. Salt concentration is shown to be a critical parameter for both the observed fluorescence intensity of ethidium intercalated in DNA, and also for the binding of positively charged polyammonium ions to DNA, effecting charge neutralisation. This charge neutralisation precedes DNA condensation, a key first step in gene therapy.
Original languageEnglish
Pages (from-to)849-859
Number of pages11
JournalJournal of Pharmaceutical and Biomedical Analysis
Volume22
Issue number5
Publication statusPublished - 2000

Fingerprint Dive into the research topics of 'Rapid and sensitive ethidium bromide fluorescence quenching assay of polyamine conjugate-DNA interactions for the analysis of lipoplex formation in gene therapy'. Together they form a unique fingerprint.

  • Cite this