Specific recognition of supercoiled plasmid DNA by affinity chromatography using the intercalator DAPP as ligand

Plasmid purification by using a new naphthalene tripodal support

“…NaCl was used to promote the binding and elution of sc pVAX1-LacZ on naphthalene tripodal support using low salt concentrations reducing the costs and the environmental impact of high salt concentrations. Moreover, NaCl increase electrostatic interactions and has proved results in purification of sc pVAX1-LacZ with DAPP support [10] and amino acids such as, L-arginine [23] and L-lysine [24].…”

“…The naphthalene tripodal ligand was designed to allow multiple non-covalent interactions (hydrophobic, electrostatic and hydrogen bonds) with pDNA, as reported for other similar ligands such as DAPP [10], berenil [11,12], L-tryptophan [9] and L-tyrosine [8]. DAPP is a groove binder, and the interaction with pDNA occurs mainly through p-p stacking and hydrophobic interactions [10]. However, DAPP is dependent of pH, and when protonated binds to pDNA through electrostatic interactions between amine groups and pDNA backbone [10].…”

“…DAPP is a groove binder, and the interaction with pDNA occurs mainly through p-p stacking and hydrophobic interactions [10]. However, DAPP is dependent of pH, and when protonated binds to pDNA through electrostatic interactions between amine groups and pDNA backbone [10]. Berenil support was also successful applied for pDNA purification and the binding preferences are similar to DAPP [12].…”

“…Surface plasmon resonance (SPR) is a technique that was used to measure interactions between the immobilized ligands, naphthalene tripodal and DAPP, on the sensor surface and sample solutions of pVAX1-LacZ isoforms that flows across the SPR surface. The DAPP-Sepharose support was used for the purification of pharmaceutical grade sc pDNA in previous studies [10,22]. Due to chemical similarities (condensed aromatic moiety) to naphthalene tripodal ligand, we chosen DAPP to immobilize it in the SPR surface for comparing the binding constants with our ligand.…”

“…Aromatic ligands are promisor molecules in sc pDNA purification due to the unique features derived from the presence of the aromatic ring [7]. Recently, several studies report the use of aromatic amino acids (L-tyrosine [8] and L-tryptophan [9]) and synthetized aromatic compounds (DAPP [10] and berenil [11,12]) for pDNA purification.…”