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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

Sigma-Aldrich

Methanol, NMR reference standard

Sigma-Aldrich

Methanol, SAJ special grade

Sigma-Aldrich

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

Sigma-Aldrich

Methanol, HPLC Plus, ≥99.9%, poly-coated bottles

Supelco

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

Imidazole, European Pharmacopoeia (EP) Reference Standard

Ondansetron impurity E, European Pharmacopoeia (EP) Reference Standard

Supelco

Residual Solvent - Acetonitrile(solution in DMSO), Pharmaceutical Secondary Standard; Certified Reference Material

Sigma-Aldrich

Imidazole, ACS reagent, ≥99% (titration)

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Methylamine solution, 40 wt. % in H₂O

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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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Methanol, anhydrous, 99.8%

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Methylamine solution, 2.0 M in methanol

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Methylamine solution, 2.0 M in THF

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Azidomethyl phenyl sulfide, 95%

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Trichloroacetic acid, ACS reagent, ≥99.0%

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Water, PCR Reagent

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Trichloroacetic acid, ≥99.0% (titration)

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Methanol, suitable for HPLC, gradient grade, 99.93%

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

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Triethylamine, SAJ first grade, ≥98.0%

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Methanol, JIS 300, ≥99.8%, for residue analysis

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Methanol, JIS special grade, ≥99.8%

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Methanol, SAJ first grade, ≥99.5%

Sigma-Aldrich

Acetonitrile solution, contains 0.05 % (v/v) trifluoroacetic acid

Pure Water Density Standard, UKAS ISO/IEC17025 and ISO Guide 34 certified, density: 0.9982 g/mL at 20 °C, density: 0.9970 g/mL at 25 °C

Sigma-Aldrich

Imidazole, for molecular biology, ≥99% (titration), free-flowing, Redi-Dri^™

Sigma-Aldrich

Imidazole, ReagentPlus^®, 99%, Redi-Dri^™, free-flowing

Sigma-Aldrich

Methanol, puriss., meets analytical specification of Ph Eur, ≥99.7% (GC)