Review Cell and Gene Therapy For Treating or Curing Diseases

Within cell and gene therapy lies a diverse range of therapeutic approaches all aimed at treating or curing disease by either blocking, altering or augmenting the expression of specific genes.

Gene Therapy
Gene therapy is the in vivo delivery of a nucleic acid sequence where multiple modalities exist, but in general, they can be grouped into three categories: 1) gene silencing/modulation 2) gene insertion/replacement and 3) gene editing. Delivery is typically addressed via encapsulation in viral vectors (ie. adeno-associated virus (AAV)), or lipid nanoparticles (LNPs), which become an integral part of the drug product and must be fully characterized as well.

Cell Therapy
Cell therapy is when the nucleic acid is delivered into cells ex vivo and these engineered cells are subsequently delivered into a patient. Both autologous and allogeneic cell therapies are being developed today.

At their core, cell and gene therapies are comprised of nucleic acids, proteins and lipids that need to be fully characterized and for which precise chemistry, manufacturing and control (CMC) processes must be established to ensure their safety and efficacy. Waters offers a broad set of tools, solutions and expertise in biomolecular separations, mass analysis and biophysical characterization to support the development and commercialization of these new and exciting modalities.

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Waters Cell and Gene Therapy Solutions

Review Cell and Gene Therapy For Treating or Curing Diseases

Nucleic Acids

Nucleic acid-based therapeutics, such as antisense oligonucleotides and RNAi therapies, routinely incorporate modified nucleotides for greater stability and are often conjugated to other molecules to effect targeted delivery. Being able to separate these molecules from a multitude of synthesis related impurities is critically important for their characterization and CMC testing, impurities analysis, and QC release. Additionally, it is critically important to confirm the nucleotide se-quence including the location of each modification within the sequence (aka: modification mapping).

To this end, Waters offers unique high-performance column chemistries and fit-for-purpose LC-MS workflows that deliver unparalleled separations performance for oligonucleotides and support everything from characterization and sequence confirmation to GMP validated QC release.

Featured Technology: BEH & ACQUITY PREMIER column technology, ACQUITY UPLC systems, BioAccord LC-MS system, XEVO G2-XS QTof, Vion & SYNAPT XS MS platforms, MassPREP standard, waters_connect, MassLynx, ProMass, Spectrum Tools

Lipid Nanoparticles

One way to deliver nucleic acid therapeutics to cells is to encapsulate them in lipid nanoparticles (LNPs). The size, composition, charge, and surface chemistry of LPNs are varied to improve delivery selectivity. ITC and DSC can both be used to interrogate the higher-order complexes formation and structure. RP chromatography is added to provide detail and GC-MS facilitates ID and detection of impurities of the raw materials.

Featured Technology: Charged Surface Hybrid™ C18 column chemistry, ACQUITY UPLC, Xevo TQ-GC, SFC, Nano DSC, Nano or Affinity ITC

Viral Vectors

The presence of the desired transgene encapsidated within a viral vector is a critical quality attribute (CQA) and because there is a charge difference between the two species, quantification and separation of empty/full viral vectors can be completed using anion exchange chromatography.

Additional elements to monitor that impact transfection efficiency, selectivity, and immunogenicity include the protein subunit ratio, post-translational modifications (PTMs), and size variants. The first two are best addressed by peptide mapping and the latter can be assessed using size exclusion chromatography.

Featured Technology: BioAccord LC-MS System, UNIFI, ACQUITY UPLC H-Class PLUS Bio System, ACQUITY UPLC with FLR Detector, RP C4 300Å or C18 300Å column, Protein Pak, XBridge Protein BEH450 SEC Columns, Empower Software with Auto Blend Plus Technology, Protein-Pak Hi Res Q Column

Bioprocess Attribute Monitoring

Metabolite monitoring is a useful process in the analysis of cell culture media whether it is for a viral vector product or cell therapy propagation of either autologous or allogeneic cells. In untargeted mapping, a researcher is given freedom to discover and after the best predictors are discovered, an automated targeted validation method with Progenesis can increase efficiency.

Featured Technology: Progenesis QI, UNIFI, MassLynx, SFC, HRMS, HILIC, Triple Quad MS/MS, Patrol, AccuTag

Genomic and Cellular Biology Automation and Preparation

Scalable, consistent identification and quantitation of nucleic acids requires both accuracy and traceability built into an automated workflow. These requirements can be achieved using the highly versatile Andrew+ pipetting robot and Pipette+ smart pipetting system, benefitting from the design and execution of protocols for RT-PCR, Next Generation Sequencing, clonal isolation, and plasmid purification. Moving into to larger complexes, cell preparations for flow cytometry and staining can also be prepared using the same automation system.

Featured Technology: Andrew+, OneLab, BeadTender, Magnet+ www.AndrewAlliance.com