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High-throughput chemical modification of oligonucleotides for systematic structure-activity relationship evaluation.

High-throughput chemical modification of oligonucleotides for systematic structure-activity relationship evaluation.

Bioconjugate chemistry (2014-11-15)

Daniel Zewge, Francis Gosselin, Denise M Kenski, Jenny Li, Vasant Jadhav, Yu Yuan, Sandhya S Nerurkar, David M Tellers, W Michael Flanagan, Ian W Davies

ABSTRACT

Chemical modification of siRNA is achieved in a high-throughput manner (96-well plate format) by copper catalyzed azide-alkyne cycloadditions. This transformation can be performed in one synthetic operation at up to four positions with complete specificity, good yield, and acceptable purity. As demonstrated here, this approach extends the current synthetic options for oligonucleotide modifications and simultaneously facilitates the systematic, rapid biological evaluation of modified siRNA.

MATERIALS

Product Number

Brand

Product Description

Supelco

Water, for TOC analysis

Supelco

Water, suitable for ion chromatography

Imidazole, European Pharmacopoeia (EP) Reference Standard

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Triethylamine, analytical standard

Ondansetron impurity E, European Pharmacopoeia (EP) Reference Standard

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Benzyl azide solution, ~0.5 M in dichloromethane, ≥95.0% (HPLC)

Supelco

Density Standard 998 kg/m³, H&D Fitzgerald Ltd. Quality

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Water, ACS reagent, for ultratrace analysis

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Imidazole, Pharmaceutical Secondary Standard; Certified Reference Material

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2,6-Lutidine, ReagentPlus^®, 98%

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Water, for cell biology, sterile ultrafiltered

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Water, tested according to Ph. Eur.

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Water, for molecular biology, sterile filtered

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Azidomethyl phenyl sulfide, 95%

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Methylamine solution, 40 wt. % in H₂O

Supelco

2,6-Lutidine, analytical standard

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Imidazole, BioUltra, for molecular biology, ≥99.5% (GC)

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Imidazole, BioUltra, ≥99.5% (GC)

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Imidazole, puriss. p.a., ≥99.5% (GC)

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Triethylamine, puriss. p.a., ≥99.5% (GC)

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Triethylamine, for amino acid analysis, ≥99.5% (GC)

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Triethylamine, BioUltra, ≥99.5% (GC)

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Triethylamine, for protein sequence analysis, ampule, ≥99.5% (GC)

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Imidazole, ReagentPlus^®, 99%

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Methylamine solution, 33 wt. % in absolute ethanol ((denatured with 1% toluene))

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Triethylamine, ≥99%

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Trichloroacetic acid, ≥99.0% (titration)

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