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

被引：149

作者：

Geall, AJ ^{[1
]}

Blagbrough, IS ^{[1
]}

机构：

[1] Univ Bath, Dept Pharm & Pharmacol, Bath BA2 7AY, Avon, England

来源：

JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS | 2000年 / 22卷 / 05期

关键词：

cholesterol polyamine carbamates; DNA binding affinity and condensation; ethidium bromide; fluorescence quenching; intercalation; lipoplexes in gene therapy;

D O I：

10.1016/S0731-7085(00)00250-8

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

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. (C) 2000 Elsevier Science B.V. All rights reserved.

引用

页码：849 / 859

页数：11

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