Recent developments in oligonucleotide research have established the potential utility of oligonucleotides as therapeutic agents, particularly in the treatment of cancer, AIDS, and other diseases. Therapeutic oligonucleotides include triplex DNA, transcription factor decoys, antisense DNA/RNA, ribozymes, PNA, 2',5'-oligoadenylates, RNAi, and CpG immune-stimulating oligomers, diagnostic oligonucleotide chips (DNA chips), and gene therapy vectors. The detection, characterization and quantification of incomplete synthesis, modified or improperly synthesized oligonucleotides is regarded as essential for demonstrating safety and efficacy of these molecules and is required by FDA and other regulatory agencies.

Solutions

Dionex offers the industry standard DNAPac^® and ProSwift^™ columns for high-resolution analysis and purification of synthetic oligonucleotides. The new bio-inert UltiMate^® 3000 Titanium analytical HPLC is designed specifically for biomolecules separation. The columns and HPLC system can be operated under denaturing conditions, such as high temperature (up to 90 °C) or high-pH eluents (up to pH 12.5). They are compatible with organic modifiers and different salts. Scaling up from analytical to semi-preparative methods is simple and cost effective.

Dionex solutions columns support:

• n – 1 resolution of synthetic oligonucleotides (up to 70-mer)
• Phosphorothioate oligonucleotides
• Linear double-stranded DNA
• Supercoiled versus nicked/linear DNA
• Linkage variants (2’-5’ versus 3’-5’)
• Tagged oligonucleotides

Separation of 1.5 μg of pd(A)_40-60 Using the DNAPac PA100 Column

Separation of Oligonucleotides by Length