Separation of synthetic oligonucleotide dithioates from monothiophosphate impurities by anion-exchange chromatography on a Mono-Q column

Xianbin Yang, Richard P. Hodge, Bruce A. Luxon, Robert Shope, David G. Gorenstein

    Research output: Contribution to journalArticlepeer-review

    21 Scopus citations

    Abstract

    A method using a strong anion-exchange liquid-chromatography column, Mono-Q, has been developed for high-resolution analysis and purification of oligonucleotide dithioates, which were synthesized by an automated, solid-phase, phosphorothioamidite chemistry. High-resolution separation of oligonucleotide phosphorodithioates from monothiophosphate impurities was obtained. High-resolution separation was also demonstrated at pH 8. The separation of oligonucleotide dithioates was found to be linearly dependent on the number of sulfurs for the same sequence length. Thiocyanate, SCN-, as eluting anion, can be used to purify oligonucleotides containing a high percentage of phosphorodithioate linkages in lower salt concentrations and provides better separation than chloride as eluting anion.

    Original languageEnglish (US)
    Pages (from-to)92-99
    Number of pages8
    JournalAnalytical Biochemistry
    Volume306
    Issue number1
    DOIs
    StatePublished - Jul 1 2002

    Keywords

    • Anion-exchange chromatography
    • Mono-Q column
    • Oligonucleotide phosphorodithioate
    • Oligonucleotide phosphoromonothioate

    ASJC Scopus subject areas

    • Biophysics
    • Biochemistry
    • Molecular Biology
    • Cell Biology

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