title: Large-scale purification of antisense oligonucleotides by high-performance membrane adsorber chromatography.
abstract: 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.
authors.affiliation: Isis Pharmaceuticals, Inc., Carlsbad, CA 92008, USA. Deshmur@war.wyeth.com
authors.forename: R R
authors.initials: RR
authors.lastname: Deshmukh
authors.forename: T N
authors.initials: TN
authors.lastname: Warner
authors.forename: F
authors.initials: F
authors.lastname: Hutchison
authors.forename: M
authors.initials: M
authors.lastname: Murphy
authors.forename: W E
authors.initials: WE
authors.lastname: Leitch
authors.forename: P
authors.initials: P
authors.lastname: De Leon
authors.forename: G S
authors.initials: GS
authors.lastname: Srivatsa
authors.forename: D L
authors.initials: DL
authors.lastname: Cole
authors.forename: Y S
authors.initials: YS
authors.lastname: Sanghvi
mesh_terms: D002852:Chromatography, Ion Exchange / Q000379:methods*; D008566:Membranes / Q000737:chemistry; D016376:Oligonucleotides, Antisense / Q000302:isolation & purification*; D013997:Time Factors
chemical_list: D016376:Oligonucleotides, Antisense