Separation of synthetic oligonucleotide dithioates from monothiophosphate impurities by anion-exchange chromatography on a mono-q column.
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.
Yang
Xianbin
X
Sealy Center for Structural Biology and Department of Human Biological Chemistry & Genetics, University of Texas Medical Branch at Galveston, TX 77555, USA.
Hodge
Richard P
RP
Luxon
Bruce A
BA
Shope
Robert
R
Gorenstein
David G
DG
D000837:Anion Exchange Resins / Q000737:chemistry*; D001483:Base Sequence; D002852:Chromatography, Ion Exchange / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D009682:Magnetic Resonance Spectroscopy; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D010710:Phosphates / Q000737:chemistry; D017553:Phosphorus Compounds / Q000737:chemistry; D012117:Resins, Synthetic; D013457:Sulfur Compounds / Q000737:chemistry; D013997:Time Factors
D000837:Anion Exchange Resins; D009838:Oligodeoxyribonucleotides; D010710:Phosphates; D017553:Phosphorus Compounds; D012117:Resins, Synthetic; D013457:Sulfur Compounds; C042855:Mono Q