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 journalArticle

19 Citations (Scopus)

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
Externally publishedYes

Fingerprint

Chromatography
Oligonucleotides
Anions
Impurities
Liquid chromatography
Sulfur
Liquid Chromatography
Purification
Chlorides
Salts
Mono Q
phosphorodithioic acid

Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Separation of synthetic oligonucleotide dithioates from monothiophosphate impurities by anion-exchange chromatography on a Mono-Q column. / Yang, Xianbin; Hodge, Richard P.; Luxon, Bruce A.; Shope, Robert; Gorenstein, David G.

In: Analytical Biochemistry, Vol. 306, No. 1, 01.07.2002, p. 92-99.

Research output: Contribution to journalArticle

Yang, Xianbin ; Hodge, Richard P. ; Luxon, Bruce A. ; Shope, Robert ; Gorenstein, David G. / Separation of synthetic oligonucleotide dithioates from monothiophosphate impurities by anion-exchange chromatography on a Mono-Q column. In: Analytical Biochemistry. 2002 ; Vol. 306, No. 1. pp. 92-99.
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