Characterisation of the binding interaction between poly(L-lysine) and DNA using the fluorescamine assay in the preparation of non-viral gene delivery vectors.
A major factor limiting the development of non-viral gene delivery systems is the poor characterisation of polyelectrolyte complexes formed between cationic polymers and DNA. The present study uses the fluorescamine reagent to improve characterisation of poly(L-lysine) (pLL)/DNA complexes post-modified with a multivalent hydrophilic polymer by determining the availability of free amino groups. The results show that the fluorescamine reagent can be used to monitor the self-assembly reaction between pLL and DNA and the degree of surface modification of the resultant complexes with a hydrophilic polymer. This experimental approach should enable the preparation of fully defined complexes whose properties can be better related to their biological activity.
Read
M L
ML
CRC Institute for Cancer Studies, University of Birmingham, Birmingham, UK.
Etrych
T
T
Ulbrich
K
K
Seymour
L W
LW
D000818:Animals; D002417:Cattle; D004247:DNA / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004996:Ethidium / Q000378:metabolism; D005450:Fluorescamine / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000378:metabolism; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D009690:Nucleic Acid Conformation; D011107:Polylysine / Q000378:metabolism*; D011108:Polymers / Q000138:chemical synthesis; D013950:Thymus Gland / Q000378:metabolism; D013997:Time Factors
D005456:Fluorescent Dyes; D011108:Polymers; D011107:Polylysine; D005450:Fluorescamine; D004247:DNA; D004996:Ethidium