Heather Stark, the Global Segment Manager at Oxford Nanopore Technologies, introduced the webinar “Enhancing Cell and Gene Therapy Success with Multi-Omic Sequencing: The Oxford Nanopore Advantage.” Start covers biopharma and explained how nanopore sequencing works. They explained that there are several end-to-end workflows for synthetic biology, including plasmid, AAV, amplicon, and mRNA IVT sequencing. Clark also listed end-to-end workflows for human research: WGS, cDNA, tumor/tumor normal, and single-cell workflows. Next, Andrea O’Hara from Azenta Life Science then presented “Using Oxford Nanopore’s Multi-Omic Sequencing to Optimize Plasmids and Vectors for Cell and Gene Therapies.”Azenta Life Sciences includes GeneWiz and several other life science companies. They presented a diagram describing the gene therapy development pipeline with three stages: discovery and identification, preclinical development, and clinical development and use. O’Hara noted that there are two ways to create an AAV plasmid. The ITR sequence is critical yet tricky to sequence. ONT Plasmid-EZ is the GeneWiz complete plasmid sequencing system. Random linearization and sequencing are then performed. The packaging components also need to be verified. Next, O’Hara explained that NGS methods for QC of viral packaging products are available. Specialized library preparations for AAV help detect contaminants such as backbone issues and host cell contaminants. O’Hara then spoke about mRNA therapeutics. The process begins with creation of a plasmid. Sequencing of the complete plasmid is necessary to verify the backbone, promoter, poly(A) tail, and region of interest. ONT can be used for sequencing the mRNA therapeutic. For clinical trials, there FDA requirements. O’Hara noted that integration site analysis (ISA) is needed for AAV gene therapy. Sequencing can be targeted or general. O’Hara ended the webinar by sharing that Azenta is used by four thousand partners and the thirty top pharma companies. This session was brief and packed with techniques used for confirmation and testing.
