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[Newsletter]
Dive into our recently published Genetic Medicine Application Notes
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Part 1: Intact RNA analysis
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Small interfering RNA (siRNA) is increasingly vital in gene therapy and cancer treatments, driving the need for advanced analytical solutions. This application note highlights how column temperature can be leveraged in chromatographic separations for siRNA duplexes, demonstrating how temperature adjustments can aid duplex sequence confirmation via intact or denatured analysis. Utilizing high resolution LC-MS Systems, the study showcases automated, compliance-ready workflows for quality control and sequence analysis.
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The use of HILIC for UV and MassSpec detection is crucial in advancing gene therapy applications, enabling precise analysis of oligonucleotide therapeutics such as siRNA duplexes, lipid-conjugated ASOs, and CRISPR sgRNAs. This application note demonstrates how HILIC-MS on an intact sgRNA for intact mass confirmation can be achieved using a benchtop, compact ToF, providing a robust and sensitive method for therapeutic RNA analysis. The study explores the application of HILIC for the separation of oligonucleotide diastereomers, duplexes, and single-stranded components, highlighting its compatibility with mass spectrometry (MS) and eliminating the need for ion-pairing agents. Key parameters, including ionic strength, temperature, and mobile phase composition, were systematically optimized using design of experiments (DoE) to improve sensitivity.
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Recent advances in LC-MS technology and informatics are enhancing mRNA critical attribute analysis. This application note outlines a workflow for mRNA sequence mapping using RapiZyme Cusatavin and RapiZyme MC1 enzymes and UPLC-QTof MS, highlighting improved sequence coverage in combination with a new informatics workflow. The study demonstrates the benefits of combining novel RNA digestion enzymes with automated software for routine, comprehensive mRNA analysis.
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The use of specific RNases is crucial in advancing gene therapy applications, enabling precise RNA sequence confirmation and modification mapping. The application note demonstrates how these RNases, combined with LC-MS and waters_connect software, provide complete sequence coverage and reproducible results for therapeutic RNA analysis. The protocol includes data processing software, such as the MAP Sequence app within waters_connect, which streamlines data analysis for routine RNA fragment identification, sequence coverage assessment, and quantification. This approach ensures simple, reproducible, and automated workflows without the need for chemical denaturants or enzyme inhibitors.
Read now →
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