Large-scale purification of antisense oligonucleotides by high-performance membrane adsorber chromatography.
Very high flux ion-exchange membranes were utilized for a novel purification of antisense oligonucleotides (20-mer). Strong anion-exchange membranes were produced by attaching polymeric ligands onto a microporous cellulosic matrix. The oligonucleotides purified were therapeutic single-stranded phosphorothioates deoxyribonucleotides. Although small-scale membrane devices (15 cm2) had similar resolution to traditional chromatographic columns; their throughputs were superior. Greater than a 1300-fold scale-up produced very similar purity and yields of the phosphorothionate product. Scale-up experiments were conducted with a 2 m2 surface area membrane module. These modules were easily capable of very high throughputs of 0.5 to 2 l/min. High purity and yields were achieved by both step and linear gradient elution.
Deshmukh
R R
RR
Isis Pharmaceuticals, Inc., Carlsbad, CA 92008, USA. Deshmur@war.wyeth.com
Warner
T N
TN
Hutchison
F
F
Murphy
M
M
Leitch
W E
WE
De Leon
P
P
Srivatsa
G S
GS
Cole
D L
DL
Sanghvi
Y S
YS
D002852:Chromatography, Ion Exchange / Q000379:methods*; D008566:Membranes / Q000737:chemistry; D016376:Oligonucleotides, Antisense / Q000302:isolation & purification*; D013997:Time Factors
D016376:Oligonucleotides, Antisense