Oligonucleotide and Nucleic Acid Standards

Oligonucleotide Specific Analogs

Analytical LC-based methods can become powerful tools for characterizing and confirming the quality of gene therapeutics, whether they be synthetic oligonucleotides, mRNA or vectored transgenes. A method has to be developed, qualified, and then checked for system suitability if it is to be appropriately implemented. Having quality reference materials on hand that are chemically similar to your analytes can make a world of difference.

Our diverse catalog of reference standards includes small interfering RNA (siRNA), lipid-conjugated antisense oligonucleotide (C16-ASO), ssDNA 20-mer, ssDNA length ladders and single guide RNA (sgRNA). These application-specific standards provide ready-to-inject references for method development, intact and MS/MS fragmentation, and system suitability testing.

Climbing the Chromatographic Ladder

Bracket your separation with 15 to 35-mer, 20 to 60-mer and 20 to 100-mer ssDNA ladders for IPRP, HILIC, and AEX analysis. Check the performance of your assays for purity, size variants, failure sequences (N/N-1) nucleotide modifications, and backbone variants. Confirm your pumps are delivering precise gradients, check the selectivity of your adsorption chromatography column, and calibrate the size-based resolving power of your separation.

Unlock SEC, AEX, and RP for Large Nucleic Acids

Nucleic acid therapeutics aren’t getting any shorter. As LC methods improve for even larger nucleic acid species, there also comes a need for new proficiency and system suitability standards. The dsDNA 50 to 1350 bp ladder is certified with wide pore SEC chromatography and can also be readily tested by anion exchange (AEX) and non-denaturing reversed-phase chromatography. The 17 double stranded DNA (dsDNA) components can be used as a reference point for size calibration, system suitability, and method troubleshooting.

Check your Extraction Efficiency

To successfully discern the Drug Metabolism and Pharmacokinetic (DMPK) properties of antisense oligonucleotides (ASO), an analyst must establish robust and efficient extraction procedures. ASO conjugation with lipid moieties enhances targeted delivery into various tissues, but this functionality has proven inherently difficult to recover from biological matrices due to solubility challenges and extreme levels of protein binding. The Waters lipid conjugated ASO LC-MS standard is conjugated with a C16 5’ moiety, and it includes 2’-MOE modifications and a phosphorothioate backbone. It is a highly discriminating reference material and can be used to evaluate the recovery of synthetic therapeutic oligonucleotides to better ensure the development of quantitative sample preparation procedures, such as the protocols provided with OligoWorks SPE.

Confirm Non-Denaturing Analyses of Duplex siRNA

Analysis of siRNA in both denaturing and non-denaturing conditions allows for full characterization of product related impurities. Both IP-RP and HILIC methods can be easily applied with both denaturing and non-denaturing conditions to help support purity and identity measurements of small interfering RNA (siRNA). Above of ~40-50 °C, most siRNA duplexes will begin dissociating into their individual sense and anti-sense single strand constituents. With an appropriately developed, non-denaturing LC assay, it becomes possible to probe duplex formation and quantify the presence of excess non-hybridized single stranded RNA (ssRNA) impurities. The Waters siRNA LC-MS Standard can be readily used to develop and qualify these assays.