Characterisation of the binding interaction between poly(L-lysine) and DNA using the fluorescamine assay in the preparation of non-viral gene delivery vectors

FEBS Lett. 1999 Nov 12;461(1-2):96-100. doi: 10.1016/s0014-5793(99)01435-0.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cattle
  • DNA / metabolism*
  • Dose-Response Relationship, Drug
  • Ethidium / metabolism
  • Fluorescamine / metabolism*
  • Fluorescent Dyes / metabolism
  • Genetic Therapy / methods*
  • Genetic Vectors / chemistry*
  • Hydrogen-Ion Concentration
  • Kinetics
  • Molecular Weight
  • Nucleic Acid Conformation
  • Polylysine / metabolism*
  • Polymers / chemical synthesis
  • Thymus Gland / metabolism
  • Time Factors

Substances

  • Fluorescent Dyes
  • Polymers
  • Polylysine
  • Fluorescamine
  • DNA
  • Ethidium