Synthetic polycation: Polynucleotide interactions determined using liquid chromatography with short monolithic columns

Abstract

LC on short monolithic columns (Convective Interaction Medium Disks) was applied to investigate several specially synthesized water soluble polycations of different charge type (primary, tertiary, quaternary amine), as well as a copolymer of neutral saccharide and cationic monomers, regarding their ability to form reversible complexes with DNA. For this purpose, two separation modes were used, namely, pseudo-affinity and cation-exchange chromatography. Synthetic polynucleotides, namely, polyriboadenylic acid (poly(rA)) and polyribocytidylic acid (poly(rC)), were used as approximate structural analogues of DNA. In first case, the hypothetical specific binding between dissolved polymers and polynucleotide (poly(rA) or poly(rC)), covalently attached to epoxy-bearing monolithic sorbent, has been studied and compared to the results obtained using cation exchange chromatography. Quantitative parameters of interactions between macromolecules were established using frontal elution method.

Anti-inflammatory effects of spermidine in lipopolysaccharide-stimulated BV2 microglial cells

“…Chemically, polyamines are cationic molecules with positive charges that enable electrostatic interactions with polyanionic macromolecules within living cells [12-14]. Several recent studies have suggested that polyamines exert multiple effects including anti-oxidant and anti-inflammatory benefits.…”

Monolithic media for applications in affinity chromatography

“…The interaction of DNA with polycations for gene delivery is still a topic of great interest 112, 113. Poly(rC)‐ and poly(rA)‐modified CIM discs have been used for studying the interactions with synthetic polycations for a potential application in gene therapy 114. Also, DNA aptamers have been used for the enrichment of proteins 109, 110.…”

Macroporous monolithic supports for affinity chromatography

“…Since AC is based on the specific interaction between the immobilized ligand and a target molecule, this type of chromatography is also used to perform a quantitative characterization of the complex formed between biomolecules and compounds with pharmaceutical interest 80. In this manner, Platonova et al used monolithic disks to study the interactions between a polynucleotide bounded in the polymeric surface and different polycations (which differ in structure and charge type) with potential application as non‐viral vectors for gene delivery 81. Two synthetic polynucleotides (poly(rA) and poly(rC)) were used to mimic the DNA structure and bound to the monolithic support via epoxy reactive groups.…”