Selectivity of arginine chromatography in promoting different interactions using synthetic oligonucleotides as model.
Arginine has been effectively used in several chromatography methodologies to improve recovery, resolution, and to suppress aggregation. Recently, arginine chromatography was used to fully separate supercoiled and open circular plasmid DNA isoforms. The specific recognition of supercoiled plasmid isoform by arginine was hypothesised to be due to the ability of arginine matrix to be involved in complex interactions that are partly dependent on the conformation of the DNA molecule. In light of these considerations a study was conducted to understand the several interactions that a DNA molecule can promote with the arginine support, in accordance with the chromatographic conditions established. Consequently, knowing the ideal conditions to promote the specific interactions, it could be possible to perform a more targeted and efficient purification. This work describes the chromatography of oligonucleotides with sizes up to 30 bases on the arginine-agarose gel. The effect of several conditions like hydrophobic character of the individual bases, molecular mass of the oligonucleotides, presence of secondary structures, temperature and elution buffer composition (salt and arginine supplemented buffer) was investigated. According to previous atomic data referent to possible interactions between amino acids and DNA nucleotides, arginine can preferentially interact with guanine by hydrogen bond, but other interactions (ionic interactions, van der Waals contacts, water mediated bonds) may also be present and become dominant depending on the conditions used. The results also revealed that the application of arginine in the elution buffer led to an effective elution of oligonucleotides from the arginine chromatographic support by a competition strategy. In general, it was suggested that the affinity interaction promoted by the arginine support is responsible for the specific recognition of particular oligonucleotide bases, involving multiple interactions.
Centro de Investigação em Ciências da Saúde, Universidade da Beira Interior, Covilhã, Portugal.
Angela
A
Sousa
Fani
F
Sousa
João A
JA
Queiroz
D001120:Arginine / Q000737:chemistry*; D002846:Chromatography, Affinity / Q000379:methods*; D004247:DNA / Q000737:chemistry; D005782:Gels / Q000737:chemistry; D006860:Hydrogen Bonding; D008024:Ligands; D008970:Molecular Weight; D009841:Oligonucleotides / Q000138:chemical synthesis / Q000737:chemistry*; D012680:Sensitivity and Specificity; D013696:Temperature
D005782:Gels; D008024:Ligands; D009841:Oligonucleotides; D004247:DNA; D001120:Arginine