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  • Oligonucleotide Separations with RP Phases

Oligonucleotide Separations with RP Phases

Feb 04 2019 Read 477 Times

Therapeutic oligos represent an important area of research in the pharmaceutical industry today. These drug candidates are typically 8-50 nucleotides long and contain single-stranded DNA or RNA. Oligonucleotides are typically analysed using ion-pairing chromatography. Oligonucleotides are easily degraded via phosphodiester-cleaving enzymes. However, with a thiophosphate modified phosphate group, the stability significantly increases. Phosphorothioate modified oligos will have a stereocenter at each modified phosphate group, leading to 2n-1 diastereomers, all with an individual retention time in the chromatography. For a 20-nucleotide long oligo, that’s 524 288 species present. The partial resolution of these species results in peak broadening, further complicating chromatographic separation. To resolve the problem, ion-pairing agents with longer alkyl chains can be used.

In a study of the influence of stationary phase and ion-pairing agent on the separation of oligos, Kromasil Phenyl showed to be a very good choice when separating oligonucleotides in general, exhibiting very sharp peaks, and good selectivity.

View these applications on www.kromasil.com/oligos.

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