Aromatic interaction chromatography : Acriflavin gel for separation of nucleotides, oligonucleotides and nucleic acids

Abstract

Using aromatic ligands anchored on insoluble polysaccharide matrices, it is possible to separate many classes of synthetic and naturally occurring substances according to their aromaticity, hydrophobicity or electrostatic character. The strength of the interactions, mainly charge-transfer interactions which occur between ligand and solute, are functions of (i) the electron donor-acceptor properties of the ligand (type, number and electron-attracting or -releasing substituents on the aromatic ring), (ii) the coupling method, (iii) the degree of substitution and (iv) the adsorption and desorption conditions of the solute sample. Different applications of this type of chromatography to the separation of nucleotides, oligonucleotides and nucleic acids are presented. With acriflavin-dextran, methods have been developed for measuring cyclic AMP in cells pre-labelled with radioactive adenine and a rapid direct assay of 3′,5′-cyclic nucleotide phosphodiesterase activity. With highly substituted acriflavin-agarose it is possible to separate a series of oligonucleotides. The low acriflavin ligand density on agarose permits the separation of single-stranded from double-stranded nucleic acids.