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Updated: Mar 9, 2026

Chemical Triphosphorylation of Oligonucleotides
Published on: June 2, 2022
Evaluation of size-exclusion chromatography for the analysis of phosphorothioate oligonucleotides
Atsuko Shimoyama1, Aki Fujisaka2, Satoshi Obika3
1Analytical Research Laboratories, Technology, Astellas Pharma Inc., 5-2-3 Tokodai, Tsukuba-Shi, Ibaraki 300-2698, Japan; Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan.
Abstract:
We evaluated size exclusion chromatography (SEC) for the detection of high-order structure of phosphorothioate oligonucleotides (PS-oligo). Because of strong interaction between PS-oligo and column packing material, peaks were broader and elution time was longer than those of the corresponding natural DNA oligonucleotides. However, single- and double-stranded structures of PS-oligo were clearly separated and discriminated, while single-stranded with high-order structures such as G-quadruplex and hairpin structure were not distinguished from each other.
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