TY - JOUR
T1 - Separation of synthetic oligonucleotide dithioates from monothiophosphate impurities by anion-exchange chromatography on a Mono-Q column
AU - Yang, Xianbin
AU - Hodge, Richard P.
AU - Luxon, Bruce A.
AU - Shope, Robert
AU - Gorenstein, David G.
N1 - Funding Information:
1This research was supported by DARPA (9624-107 FP), NIH (AI27744), NIEHS (ES06676), the Welch Foundation (H-1296), and the Sealy and Smith Foundation.
PY - 2002/7/1
Y1 - 2002/7/1
N2 - 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.
AB - 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.
KW - Anion-exchange chromatography
KW - Mono-Q column
KW - Oligonucleotide phosphorodithioate
KW - Oligonucleotide phosphoromonothioate
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U2 - 10.1006/abio.2001.5694
DO - 10.1006/abio.2001.5694
M3 - Article
C2 - 12069419
AN - SCOPUS:0036629203
SN - 0003-2697
VL - 306
SP - 92
EP - 99
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 1
ER -