Nucleic Acid Solutions

Nucleic Acid Solutions

Comprehensively obtain purity, identity, potency, and safety indicating measurements for long-chain nucleic acids (mRNA, ssDNA, DNA) using robust, compliance-ready, easy-to-implement tools and techniques from Waters.

Comprehensively obtain purity, identity, potency, and safety indicating measurements for long-chain nucleic acids (mRNA, ssDNA, DNA) 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 in length from 500 to 10,000 nucleotides or more. Confirming their identity and purity is critically important to ensure 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.

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

Applications

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

Application Notes

Application Notes

The concentration of a nucleic acid therapeutic must be accurately determined to ensure the proper dosing and consistent therapeutic outcomes across different batches of drug product. Reversed-phase chromatography and anion exchange separations can be effectively applied for this purpose.

The concentration of a nucleic acid therapeutic must be accurately determined to ensure the proper dosing and consistent therapeutic outcomes across different batches of drug product. Reversed-phase chromatography and anion exchange separations can be effectively applied for this purpose.

Application Notes

Application Notes

5’ cap analysis, 3’ poly A tail analysis, and oligonucleotide mapping for sequence and modification confirmation are crucial to verify the integrity, translation efficiency, and stability of mRNA. These attributes are directly tied to clinical performance and safety. Waters advanced analytics integrate the use of precise digestion enzymes and routine software workflows to enhance RNA analysis, ensuring comprehensive and reliable mRNA evaluation.

5’ cap analysis, 3’ poly A tail analysis, and oligonucleotide mapping for sequence and modification confirmation are crucial to verify the integrity, translation efficiency, and stability of mRNA. These attributes are directly tied to clinical performance and safety. Waters advanced analytics integrate the use of precise digestion enzymes and routine software workflows to enhance RNA analysis, ensuring comprehensive and reliable mRNA evaluation.

Application Notes

Application Notes

Recombinantly expressed plasmids are essential raw materials for viral vector production and in-vitro transcription (IVT) reactions used to manufacture mRNA. For IVT applications, confirming complete and accurate linearization of the plasmid is critical, as residual supercoiled or open circular forms can lead to premature transcription termination and heterogeneous mRNA products. Analyzing plasmid topology for linearization efficiency prior to downstream bioprocessing ensures the integrity and efficacy of the resulting mRNA drug substance.

Recombinantly expressed plasmids are essential raw materials for viral vector production and in-vitro transcription (IVT) reactions used to manufacture mRNA. For IVT applications, confirming complete and accurate linearization of the plasmid is critical, as residual supercoiled or open circular forms can lead to premature transcription termination and heterogeneous mRNA products. Analyzing plasmid topology for linearization efficiency prior to downstream bioprocessing ensures the integrity and efficacy of the resulting mRNA drug substance.

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.
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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 Solutions

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 critical quality attribute confirmation and monitoring throughout development and manufacture

Efficient RNA critical quality attribute confirmation and monitoring throughout development and manufacture
BioAccord LC-MS System

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 applications for compliance-ready data analysis and reporting.

  • Integrity and Aggregate Analysis
  • Sequence and Modification Confirmation

Exceptional performance, without compromise for RNA attribute characterization and monitoring

Exceptional performance, without compromise for RNA attribute characterization and monitoring
Xevo MRT Mass Spectrometer

Confidently identify and monitor RNA quality attributes using the Xevo MRT on waters_connect. Sub-ppm mass accuracy, efficient fragmentation, high mass resolution, and powerful quantitative capabilities enhance both accuracy and lab productivity.

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

Complete high-resolution coverage for confident RNA characterization and monitoring

Complete high-resolution coverage for confident RNA characterization and monitoring
Xevo G3 QTof

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
DAWN® MALS Detector

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

Small Volume Gene Therapy Particle Characterization

Small Volume Gene Therapy Particle Characterization
Aura GT from Halo Labs

Gain unprecedented insight into gene therapy development with Aura GT. Just 5 µL of sample is all you need to evaluate aggregation and nucleic acid leakage across a range of gene delivery modalities—including AAV, LVV, and LNP—so you can identify stability issues early and ensure product quality.

  • Integrity and Aggregate Analysis

Compliance-ready, networkable applications-based software to streamline LC-MS data acquisition, processing, and reporting.

Purposefully designed RNA informatics workflows

Purposefully designed RNA informatics workflows
waters_connect for biopharmaceuticals

Accelerate the journey from sample analysis to decision-making with compliance-ready MAP Sequence, CONFIRM Sequence and INTACT Mass Apps for comprehensive nucleic acid product understanding.

  • Integrity and Aggregate Analysis
  • Sequence and Modification Confirmation

Columns and analytical standards that deliver leading-edge LC and LC-MS performance for nucleic acids

RPLC columns for exceptional resolution, reproducibility, and stability

RPLC columns for exceptional resolution, reproducibility, and stability
MaxPeak Premier BEH C18 Columns for Oligonucleotide Analysis

Obtain exceptional ion pair reversed phase (IP-RP) resolution, sensitivity, and stability with Waters MaxPeak Premier Oligonucleotide BEH C18 Columns that are QC-tested with Waters MassPrep Oligonucleotide Standard to ensure lot-to-lot reproducibility.

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  • Concentration Determining Assays
  • Sequence and Modification Confirmation

High-efficiency inert wide-pore SEC Columns

High-efficiency inert wide-pore SEC Columns
GTxResolve SEC Columns

Achieve intact separation of long-chain nucleic acids, including mRNA and their fully packaged drug product, with Waters GTxResolve SEC 450 Å, 1000 Å, and 2000 Å columns that separate nucleic acids and assembled drug products based on size and shape, enabling payload quantification, product integrity, and purity analyses.

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

Orthogonal selectivities and ion pairing free chromatography

Orthogonal selectivities and ion pairing free chromatography
Gene-based Therapeutics (GTx) Columns

Obtain orthogonal selectivity and ion pairing free LC options with Waters Gene-based Therapeutics (GTx) Columns, including weak, and strong anion exchange columns and amide-bonded BEH particle HILIC columns for an array of new method options.

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

Perform system suitability and benchmark methods with confidence

Perform system suitability and benchmark methods with confidence
Oligonucleotide and Nucleic Acid Standards

Eliminate the need to prepare complex mixtures with Waters prepackaged lot-certified Oligonucleotide and Nucleic Acid Standards that deliver ease-of-use and peace-of-mind when running or troubleshooting important separation-based assays.

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

RNA Digestion with Unique Cleavage Specificities

RNA Digestion with Unique Cleavage Specificities
RapiZyme RNases

Observe 3x increased sequence coverage and longer RNA digestion products with unique masses for LC-MS analysis. RapiZyme RNases offer complementary cleavage properties and overlapping digest products for improved confidence in identification.

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  • Sequence and Modification Confirmation

For Long Chain Nucleic Acids

For Long Chain Nucleic Acids
Slalom Chromatography

Avoid running gels and realize tremendous time savings/greater efficiency when confirming plasmid length and determining linearization efficiency - a game changer for In Vitro Transcription (IVT) labs.

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  • Plasmid Topology and Purity Analysis

 Your success is just a click away

 Your success is just a click away
Waters Global Services

Optimize your laboratory's productivity and success with Waters Global Services to maintain peak system performance, minimize down time, address application challenges, and support stringent compliance requirements.

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

Make science more accessible with Waters Capital

Make science more accessible with Waters Capital
Payment Solutions

Maximize resources and minimize risk with payment options from Waters Capital, including upgrading aging equipment, getting customized support, and bundling services into one monthly payment.

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

The data speaks for itself

The data speaks for itself
rapizyme-cusativin-digest-of-sgRNA

LC-UV-MS analysis of the HPRT sgRNA digested with recombinant RapiZyme Cusativin. Three independent recombinant enzyme batches of RapiZyme Cusativin were used to digest HPRT sgRNA, and the three UV chromatograms (black, blue, and red traces) are overlayed on top of each other, indicating highly reproducible digestion behavior.

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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).

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Characterization of different forms of 5’-capped oligonucleotides in the RNase MC1 digest of mRNA 1 including cap (a), unmethylated cap (b), and uncapped (c).

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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
Download
  • eBook

eBook: Characterizing LNP mRNA Consumables

eBook: Characterizing LNP mRNA Consumables
Download
  • 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
Download
  • 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
Watch Webinar
  • 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
Watch Webinar
  • Webinar

New Insights on Anion Exchange and Size Exclusion of Nucleic Acids

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

Mapping RNA: An LC-MS Exploration, BioSpring

Mapping RNA: An LC-MS Exploration, BioSpring
Watch Webinar
  • Webinar

Cracking the Code: Analyzing mRNA and sgRNA with Advanced Enzymes, Mass Spectrometry, and Informatics

Cracking the Code: Analyzing mRNA and sgRNA with Advanced Enzymes, Mass Spectrometry, and Informatics
Watch Webinar
  • Infographic

Attribute Monitoring Workflows for Nucleic Acid Therapeutics

Attribute Monitoring Workflows for Nucleic Acid Therapeutics
Download

Learn more about Nucleic Acid Solutions.

Learn more about Nucleic Acid Solutions.
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