期刊
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
卷 22, 期 5, 页码 849-859出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/S0731-7085(00)00250-8
关键词
cholesterol polyamine carbamates; DNA binding affinity and condensation; ethidium bromide; fluorescence quenching; intercalation; lipoplexes in gene therapy
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.
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