Ja. Gerstner et al., GRAM-SCALE PURIFICATION OF PHOSPHOROTHIOATE OLIGONUCLEOTIDES USING ION-EXCHANGE DISPLACEMENT CHROMATOGRAPHY, Nucleic acids research, 23(12), 1995, pp. 2292-2299
The purification of oligonucleotides by ion-exchange displacement chro
matography is demonstrated on the gram-scale. Using a 50 mmD x 100 mmL
(203 ml) column operated in the displacement mode, 1.2 g of a 24mer p
hosphorothioate oligonucleotide was purified. Product yield for this s
eparation was 70% (780 mg) at a purity of 96.4% and the mass balance r
ecovery of all oligonucleotide was 97.5%. The displacement purificatio
n of four additional phosphorothioate oligonucleotides ranging in leng
th from 18 to 25 bases is also demonstrated on the semi-preparative (1
0-50 mg) scale. All of these oligonucleotides were purified using simi
lar displacement conditions and typical results were 60% yield at 96%
purity. The displacement portion of these separations required <15 min
and total cycle time including equilibration, feed loading and regene
ration can be performed in under 30 min. These results seem to indicat
e that displacement chromatography may be amenable to generalizations
in separation protocol that would greatly reduce the effort required t
o obtain an optimized purification scheme for moderately long oligonuc
leotides.