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CMC for Gene Therapy


On-Demand Training







Course Module 1: USP Standards and Best Practices for Gene Therapy

Jim Richardson Ph.D.

Module Description:

Gene therapies offer tremendous promise to address human disease but their complexity and diversity present unique challenges to those seeking to standardize materials and methods. USP is engaging with stakeholders to identify and develop documentary and physical standards to support gene therapy. This presentation will provide an overview of existing USP documentary and physical standards as well as an update on standards under development.

Course Module 2: Considerations for Nucleic Acids and Enzymes in Gene Therapy: Evaluating the Impact of How Materials Are Used On Design, Characterization and Specifications

James Brown Ph.D.

Module Description:

Considerations for nucleic acids and enzymes in gene therapy: evaluating the impact of how materials are used on design, characterization and specifications Recent advances in gene therapy have made treatments based on reprogramming cells to treat disease a reality. Researchers are developing a wide array of modalities to address an ever-growing list of diseases. Viral vectors are addressing monogenic defects with protein replacements, delivering therapeutic proteins, and reprogramming T cells to target cancer. Nucleic acids and gene editing nucleases are being used to manipulate various stem cells. Genetic payloads are being developed for individual patients based on the unique proteins and peptides in a tumor, resulting in a truly personalized therapy. Each of these approaches needs a variety of nucleic acids and enzymes to achieve the therapeutic benefit. How each of these are used, their proximity to the patient, the route of administration, and other factors impact the requirements of these materials. We will review examples of how raw materials are used in these therapies and how best to evaluate the design, characterization and specifications in the context of their use. We will present some of the key requirements that drive cost and timeline with a goal to optimize production and ensure the safety and efficacy of the product. We will discuss the spectrum of needs that must be addressed, from off-the-shelf products to enable large scale production of a single therapy to “off-the-shelf production methods” to enable small scale production of many individual therapies.

Course Module 3: Current Good Manufacturing Practices (CGMP) Compliance Issues for Gene Therapy Products

Heath Coats, BS, MS

Module Description:

Phase specific GMPS – Phase I Chain of Custody/Chain of Identity GMPs to ensure patient safety – Phase II and III – Phase 2 and 3 manufacturing will continue to be subject to parts 210 and 211. Raw material appropriate quality and grade – vendor and material qualification If contract manufacturer used, adequate management and oversight of CMO – Appropriate experience with product type by CMO – Appropriate KSA (Knowledge, Skill, Ability) of sponsor personnel to adequately oversee CMO – Appropriate segregation of different products if multi-product facility Dedicated/Shared/Single use equipment Understanding boundaries of shared/dedicated equipment PCR use for carryover assessment – CMO may not disclose all products manufactured due to proprietary information – reference a contract manufacturing facility’s Master Files for submissions CBER Drug Master File Guidance information https://www.fda.gov/vaccines-blood-biologics/new-drug-applicationnda-process-cber/drug-master-files-cber-regulated-products Drug Master Files Guidance for Industry (DRAFT) https://www.fda.gov/media/131861/download – Appropriate viral vector containment mitigation strategies – Appropriate review (and approval) of CMO documentation – sponsor review of CMO final product Quality agreements with suppliers, CMOs, and CTLs ALCOA data integrity Capacity throughput studies if Gene Therapy product will be introduced into patients cells then administered to patient Example 483 items

Course Module 4: Viral Vector Manufacturing and Control Strategies 

Richard Snyder, Ph.D.

Module Description:

Viral gene transfer vector manufacturing for in vivo and ex vivo applications has largely been in support of early phase clinical trials, but as product candidates move to later development stages, demand is rapidly increasing for commercial grade vectors at a variety of scales. Decisions regarding vector design, manufacturing platform, product configuration, process control, and regulatory strategy have an impact on timelines and resources, raw materials sourcing, and analytical testing. Developing a strategy that supports an efficient path to commercialization while reducing risk helps to bring these cutting edge cell and gene therapies to patients in need.

Course Module 5: Developing a Potency Assay for GeneRide Products: an AAV-based Genome Editing

Lauren Drouin, Ph.D.

Module Description:

The GeneRide platform utilizes the natural process of homologous recombination to achieve targeted genome editing without the use of exogenous nucleases or promoters. Developing a potency assay for this unique technology poses additional challenges over those for canonical gene therapy products, including the requirement of a highly sensitive detection method to measure low levels of genome integration. The first GeneRide candidate, LB-001, is currently under clinical development for the treatment of methylmalonic acidemia. LB-001 targets site-specific integration into the albumin locus to allow the gene of interest (MMUT) to be expressed concomitantly with albumin. In order to control LB-001 product activity and assess lot-to-lot consistency, a cell-based assay was developed to measure fused mRNA expression as a surrogate of biological activity. The assay was developed in a cell line that naturally expresses albumin and can thus drive expression of the MMUT gene upon site-specific integration. Fused mRNA is quantified using primers overlapping the host genome and the transgene DNA to ensure that only the integrated product is detected. The results show that AAV-driven homologous recombination is reproducible in vitro, which allows for the qualification of assay control material. The method was tested for linearity, repeatability, and specificity. Examination of the assay data demonstrates that this method is suitable for assessing the relative potency of integrating GeneRide vectors.

Course Module 6: Charge Detection Mass Spectrometry for Characterization of AAV Capsids

Benjamin E Draper, Ph.D.

Module Description:

• Charge Detection Mass Spectrometry (CDMS) is capable of making mass and charge measurements of AAV vectors.

• CDMS quantifies empty/partial/full ratios, impurities and high mass aggregates of AAV formulations.

• CDMS measurements are fast, sensitive, and comprehensive allowing measurement of low concentration and heterogeneous samples.

• CDMS can characterize AAV vectors from early to late stage development.

Course Module 7: Analytical Methodologies for Analysis of AAV Viral Vector and Nucleic Acids for Gene Therapy

Sahana Mollah, Ph.D.

Module Description:

Fast growth in cell and Gene Therapy industry has generated an urgent need for fast and robust analytics for characterization and impurity determination for both viral vectors and various types of nucleic acid biotherapeutics such as oligonucleotides, plasmid DNA, small RNA, mRNA and double stranded DNA. Although there are efforts to develop analytical assays for characterization of the nucleic acids and vectors, there are still limitations and drawbacks to some of these analysis. In addition, the different workflows are done on different platforms. Capillary electrophoresis is a widely used technique for analysis of protein and nucleic based therapeutics. Here, we present results for multiple key Gene Therapy analytical workflows performed using capillary electrophoresis (CE) based platform. These methods not only provide characterization of various nucleic acid and vectors, but also analysis of impurities in raw material and drug products. Analytical methodologies included in this presentation are i) fast, easy, orthogonal method for AAV capsid purity analysis (VP1, VP2 and VP3) with ultra-high sensitivity using capillary electrophoresis sodium dodecyl sulfate (CE-SDS) with laser induced fluorescence detection (LIF), ii) method for analysis of plasmid purity and stability using capillary gel electrophoresis (CGE) with LIF detection, iii) sizing and purity analysis of transgenes with CGE-LIF and iv) purity analysis of single guide RNA (sgRNA) and cas9 mRNA for CRISPR analysis with CGE-LIF.

Course Module 8: Characterize and Quantify AAV by DLS and SEC-MALS

Sophia Kenrick, Ph.D.

Module Description:

To ensure the safety and efficacy of viral vector-based drug products, robust and reliable characterization tools are essential throughout the production and manufacturing process. In this presentation, we discuss a size exclusion chromatography (SEC) method coupled with UV, multi-angle light scattering (MALS), and differential refractive index (dRI) detectors to measure the following three important AAV quality attributes (QAs): total number of viral capsid particles, relative capsid content, and percentage of monomer or aggregates. In addition, orthogonal screening and characterization with dynamic light scattering (DLS) and field flow fractionation (FFF) will be compared.

Course Module 9: Update from the Standards Coordinating Body in Support of Gene Therapy

Catherine Zander, Ph.D.

Course Module 10: Global Harmonization and Standards for Regenerative Medicines Therapy

Judy Arcidiacono, M.S.


Date: [course_completion_date]

Couse Duration (hh:mm:ss): 04:52:56