A resurgence in the development of oligonucleotide therapeutics has led to burgeoning pipelines across the industry. Working at the level of gene transcription and translation, these short nucleic acid drugs hold great promise for many inherited disorders and common ailments alike.
Waters LC-MS tools and techniques play a critically important role in confirming the identity and purity of these drugs, as well as quantifying their presence in biofluids.
Case Study: BioSpring Implements Waters LC-MS Workflows for Oligonucleotides to Expand Existing Portfolio
MaxPeak Premier columns and systems eliminate the unpredictability of analyte losses due to metal interactions, which can be especially pronounced with negatively charged oligonucleotides.
Simplify high-performance LC-MS biopharmaceutical analysis for every user with Waters BioAccord LC-MS System with ACQUITY Premier, an integrated, compliance-ready platform that can be deployed from discovery to GxP labs.
Accelerate characterization of synthetic oligonucleotides with the Xevo G3 QTof high-performance HRMS benchtop system that delivers accurate qualitative and quantitative results.
Expand the scope of ultimate sensitivity analysis with Waters Xevo TQ Absolute Triple Quadrupole Mass Spectrometry that delivers unprecedented levels of reliability, reproducibility, and performance for your oligonucleotide applications.
Testing the inertness of the ACQUITY Premier UPLC System and ACQUITY Premier OST Column using the 21-nt oligomer: UV chromatogram of the lowest detectable concentration (0.5 nM, or 5 femtomoles of the 21-mer oligonucleotide loaded on-column) compared against the preceding blank injection.
Calibration curves of GEM91 using a stainless-steel column and conventional system (red), an ACQUITY Premier Column and conventional system (blue), and an ACQUITY Premier Column and System (purple). The different slopes indicate distinct differences in recovery using the columns and systems tested.
TUV and TIC chromatograms showing the performance of the BioAccord System when the 1 μM OST standard was diluted 100 times (100 fmoles sample loading). All five major oligos are clearly detected in both the UV and TIC chromatograms, indicating that the LC-MS system and method has adequate sensitivity for detecting low-level oligo impurities.
LC-MS/MS full ladder sequence confirmation for the ssRNA sequence of 5'-UCGUCAAGCGAUUACAAGGTT-3' was achieved. The experiment was conducted using the M4- peak as the precursor ion. The fragmentation peak assignments were done by matching against the theoretical masses using simple Excel file calculations. MaxEnt3 was used for charge deconvolution and deisotoping.
