Nucleic Acid Solutions

Nucleic Acid Solutions

Comprehensively obtain purity, identity, potency, and safety-indicating long-chain nucleic acid measurements using robust, compliance-ready, easy-to-implement tools and techniques from Waters.

Comprehensively obtain purity, identity, potency, and safety-indicating long-chain nucleic acid measurements using robust, compliance-ready, easy-to-implement tools and techniques from Waters.

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Overview

Long nucleic acids / transgene sequences in cell and gene therapies range from 1,000 to 10,000+ nucleotides in length. Confirming their identity and purity is critically important to ensuring the safety and efficacy of these new modalities. Waters chromatography columns and systems combined with optical detection and/or mass spectrometry provide powerful capabilities for confirming the physicochemical properties of these nucleic acids and of their fully assembled drug product when packaged into lipid nanoparticles (LNPs), viral vectors (e.g. AAV), or virus-like-particles (VLPs) for cell delivery.


Applications

To ensure the stability, efficacy, and safety of cell and gene therapies, it is critically important to measure the integrity, heterogeneity, and purity of their nucleic acid payloads, whether in the form of mRNA, ssDNA, or dsDNA. Chromatography combined with light scattering and/or mass spectrometry is a powerful technique for measuring these critical quality attributes.

To ensure the stability, efficacy, and safety of cell and gene therapies, it is critically important to measure the integrity, heterogeneity, and purity of their nucleic acid payloads, whether in the form of mRNA, ssDNA, or dsDNA. Chromatography combined with light scattering and/or mass spectrometry is a powerful technique for measuring these critical quality attributes.


Application Notes

Application Notes


On-Demand Webinar: LC-MS CQA Analysis of RNA Therapeutics

In this webinar, we present analytical workflows for RNA CQA analysis with data acquired on our compliance-ready waters_connect platform. Demonstrated applications include characterization of both sgRNA (ID and structural fidelity) and mRNA (ID, structural fidelity, 5’ capping, 3’ poly-A tail) molecules.

In this webinar, we present analytical workflows for RNA CQA analysis with data acquired on our compliance-ready waters_connect platform. Demonstrated applications include characterization of both sgRNA (ID and structural fidelity) and mRNA (ID, structural fidelity, 5’ capping, 3’ poly-A tail) molecules.


Elearning concept background

Solutions


Waters LC and LC-MS systems offer leading separations performance and sensitivity for nucleic acid analysis.

Ultra-low adsorption for maximum sensitivity and reproducibility

Ultra-low adsorption for maximum sensitivity and reproducibility

MaxPeak Premier columns and systems eliminate the unpredictability of analyte losses due to metal interactions, which can be especially pronounced with negatively charged nucleic acids.

  • Integrity and Aggregate Analysis
  • Concentration Determining Assays
  • Sequence and Modification Confirmation
  • Plasmid Topology and Purity Analysis

Efficient RNA product attribute monitoring throughout development and manufacture

Efficient RNA product attribute monitoring throughout development and manufacture

Enable routine LC-MS biopharmaceutical analyses in every lab with the Waters BioAccord LC-MS System, combining the confidence of accurate mass data and waters_connect software for compliance-ready data analysis and reporting.

  • Integrity and Aggregate Analysis
  • Sequence and Modification Confirmation

Complete high-resolution coverage for confident RNA characterization and monitoring

Complete high-resolution coverage for confident RNA characterization and monitoring

Accelerate characterization  of nucleic acids with the Xevo G3 QTof high-performance HRMS benchtop system, and translate that knowledge to efficient monitoring assays, delivering accurate qualitative and quantitative results with data sharing across waters_connect networks.

  • Integrity and Aggregate Analysis
  • Sequence and Modification Confirmation

Get insight on your nucleic acid stability

Get insight on your nucleic acid stability

Obtain simultaneous measurement of size, molecular weight, distribution, and morphology with the flexible Wyatt DAWN MALS Detector coupled with Waters LC systems, including the Arc Premier System, applicable for nucleic acids, LNPs, and conjugated proteins.

  • Integrity and Aggregate Analysis
  • Concentration Determining Assays
  • Plasmid Topology and Purity Analysis


The data speaks for itself

The data speaks for itself
The most abundant peak in the Fluc Beta mRNA poly(A) tail grouping of peaks corresponds to a 128 nt long species Separation of 100 nt oligo(A) synthetic RNA oligonucleotide standard (black chromatogram), RNase T1 digested EPO mRNA (blue chromatogram) and RNase T1 digested Fluc Beta mRNA (red chromatogram) with an IP RP LC method using an ACQUITY Premier Oligonucleotide BEH 300 Å 1.7 µm Column. The most abundant peak amidst the EPO mRNA poly(A) tail series of peaks corresponds to a 124 nt long species. The most abundant peak in the Fluc Beta mRNA poly(A) tail grouping of peaks corresponds to a 128 nt long species.
The effect of temperature on the chromatographic profile (selectivity) and carryover The effect of temperature on the chromatographic profile (selectivity) and carryover. Column: Gen-Pak FAX 100 x 4.6 mm, 2.5 µm Column, mobile phase A: 25 mM TRIS, pH=7.6, mobile phase B:25 mM TRIS, pH=7.6 + 2 M NaBr, F=0.6 mL/min, gradient: 12–35% B in 15 min (shallow gradient), modulator solvent plug: mobile phase B. Bracketing injection: 1 µL modulator pre-plug + 2 µL sample + 1 µL modulator post-plug. Temperature: ambient (left), 35 °C (middle) and 45 °C (right).
The software presents confirmed fragment ions on a Dot-Map to quickly assess the sequence coverage (A) Digest products at position 623–631(ACAUCCUCGp) and 551–559 (UCACCAUCGp) are predicted and assigned to the same RT peak in the TIC from Gaye et al. It is not possible to determine the correct assignment using intact mass information. (B) MSE data from the same injection can be used to elucidate the correct sequence for this assignment. Using the waters_connect CONFIRM Sequence application, high energy fragment ions are predicted for each sequence, and matched to isotope clusters of the integrated raw data via a bespoke algorithm. The software presents confirmed fragment ions on a Dot-Map to quickly assess the sequence coverage.
Effect of pH and temperature on the ΦX174 plasmid isoform separation Effect of pH and temperature on the ΦX174 plasmid isoform separation.

Webinars and Resources


  • Brochure

Routine Attribute Monitoring Of RNA: Enabled Decisions for Consistent Product Quality

Routine Attribute Monitoring Of RNA: Enabled Decisions for Consistent Product Quality
  • eBook

eBook: Characterizing LNP mRNA Consumables

eBook: Characterizing LNP mRNA Consumables
  • Application Notebook

LC-MS Analysis Workflows for RNA Therapeutics to Ensure Product Quality and Process Consistency

LC-MS Analysis Workflows for RNA Therapeutics to Ensure Product Quality and Process Consistency
  • Poster

Characterization of Intact mRNA Using Ion Pair-Reversed Phase-Time of Flight-MS, Size Exclusion Chromatography-Multi Angle Light Scattering, and Charge Detection Mass Spectrometry

Characterization of Intact mRNA Using Ion Pair-Reversed Phase-Time of Flight-MS, Size Exclusion Chromatography-Multi Angle Light Scattering, and Charge Detection Mass Spectrometry
  • Webinar

LC-MS for RNA Therapeutics to Ensure Product Quality and Process Consistency

LC-MS for RNA Therapeutics to Ensure Product Quality and Process Consistency
  • Webinar

Unveiling CQA Insights for mRNA, LNPs and Viral Vectors by SEC and MALS with Novel 1000 Å Ultrawide Pore Particle Columns

Unveiling CQA Insights for mRNA, LNPs and Viral Vectors by SEC and MALS with Novel 1000 Å Ultrawide Pore Particle Columns
  • Webinar

New Insights on Anion Exchange and Size Exclusion of Nucleic Acids

New Insights on Anion Exchange and Size Exclusion of Nucleic Acids
  • Infographic

Attribute Monitoring Workflows for Nucleic Acid Therapeutics

Attribute Monitoring Workflows for Nucleic Acid Therapeutics

Related


Learn more about Nucleic Acid Solutions.

Learn more about Nucleic Acid Solutions.

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