Our offering

Genome engineering of the GOI (gene of interest) cassette is a dynamic process to ensure efficient gene expression, therapeutic efficacy, and viral vector manufacturability. We employ the following key strategies and considerations to optimize the GOI cassette:

• Optimize ITR sequences
• Promoter selection and possibility to include enhancer or regulatory elements
• Identify cryptic splice sites, cryptic poly A sites
• Intron inclusion to stabilize mRNA
• Codon optimization to improve translation efficiency
• CpG island optimization
• Payload corrections
• Reverse package mitigation to avoid DNA impurities in viral vector production
• Prioritize safety by avoiding or minimizing the use of immunogenic sequences

Our manufacturing platform is based on transient transfection of HEK293 cells in suspension, and customized state-of-the-art upstream and downstream processes.

We provide our partners:

• Technology transfer
• Lab-scale viral vector manufacturing
• Implementation of scalable single-use bioreactors technologies at 2-liter scale, enabling collection of initial yield and purity data specific to the construct. Process development needs are identified before future scale-up to pilot 50-liter scale
• Process design and selection of fit-for-purpose upstream and downstream processing technologies
• Characterization and optimization of upstream and downstream processes
• Formulation of drug product
• Scale-up development in 50-liter bioreactors based on previous stages. Material supplied can be used to complete research and GLP preclinical studies
• Assay implementation and quality control testing required for batch release

A DINAMIQS project manager and cross-functional team will help you meet your objectives, providing you with high quality vectors while minimizing process changes to de-risk and accelerate timelines.

We provide valuable input in the design and development of customizable GMP-compatible processes. Acting as a single service partner, we develop scale-up strategies to ensure a seamless transition between the different phases of your project, from early development to the manufacturing of GMP material for Phase I/II clinical studies. We ensure your process is robust, scalable, cost-effective and reproducible.

Our upstream development capacities integrate:

• Media and feed optimization for batch production
• Chemically defined medium, animal component-free
• Single-use technologies
• High viral titer, suspension cell line
• High viable cell density
• Adenovirus-free, triple plasmid transfection
• Optimized cell lysis
• DNA digestion with GMP-compatible endonuclease
• Scalable harvest and clarification operation

Viral vector purification is critical to ensure a high-level of product safety, including purity and potency. Our downstream development capacities integrate:

• Bioprocess filtrations (normo- or tangential flow filtration)
• Column and membrane chromatography (affinity, ion exchange, hydrophobic interaction, desalting, multimodal)
• Density gradient ultracentrifugation using Iodixanol and/or CsCl
• Design and implementation of single-use technologies
• Final filtrations with formulation and sterile filtration
• Customized fill and finish

Analytical characterization and quality control of rAAV vectors is crucial at every stage of development and manufacturing. Our robust, accurate and reproducible analytical methods accelerate process development and ensure the quality of our manufactured vector products.

We offer a full analytical service package tailored to your gene therapy product, including:

Strength and quantification

• PCR-based vector quantification: qPCR – ddPCR
• We perform ITR-based viral vector quantification and offer method development and implementation for your gene of interest sequences for quantification.
• Total capsid ELISA for all common serotypes

Potency vector transduction and infectivity

• Infectivity (TCID50 method)
• Transduction by FACS analysis (applicable to fluorescent reporter vectors)
• Expression of transgene mRNA (RT-qPCR)
• Development of customized cell-based potency assays

Purity, identity and characterization

• Protein purity and identity by SDS-PAGE/Western blotting
• Capsid protein identity by SYPRO Orange Differential Scanning Fluorimetry (protein melting temperature analysis)
• Genome integrity by alkaline gel electrophoresis
• Vector sequence identity and genome heterogeneity by long-read sequencing and bioinformatics analysis
• Particle size distribution/aggregation by dynamic light scattering (DLS)
• Empty and full viral vector particle ratios by mass photometry, qPCR/ELISA and SLS-UV/VIS

Impurities

• HEK 293 host-cell protein ELISA
• Bacterial endotoxin test (LAL kinetic turbidimetric method)
• Residual host-cell and plasmid DNA (qPCR-based quantification)

Our AAV5, AAV8 and AAV9 vectors, available as both full and empty particles, are designed to meet your needs for reliable performance and exceptional results. Use full particles for serotype screening or empty particles for immunoassays like measuring neutralizing antibodies. Additional serotypes are being produced and will soon be available. Our vectors are manufactured with materials and processes that are directly scalable to GMP, enabling a faster transition from proof of concept to IND-enabling studies.