Identity Confirmation

What is Identity confirmation and why is it important?

Identity testing is a critical component to release vector products, including viral and non-viral cell and gene therapies, as well as vaccine and oncolytic applications. Determination of the nucleic acid sequence of the vector and gene of interest is a regulatory requirement, with full nucleic acid sequence provision for vectors under 40kb. Therefore, direct sequencing is a standard and well-accepted approach.

Sanger sequencing has traditionally been employed for nucleic acid testing and is well accepted by regulators. However, an NGS approach provides several advantages over Sanger sequencing:

• Identify ultra-rare genetic variants <<5% (~20% variant detection limit for Sanger).
• More cost and time efficient over a sequential Sanger sequencing.
• Full coverage can be challenging with Sanger sequencing

As there is a regulatory expectation for appropriate characterization of critical raw materials, an aligned nucleic acid analysis strategy should also be employed for any plasmid input materials. This mitigates the risk that non-clonal variation within the manufacturing plasmids will affect the quality of the final product.

NGS screening offers several advantages, including the ability to detect ultra-rare genetic variants and providing more comprehensive coverage than traditional methods. This makes NGS a superior choice for nucleic acid testing in the context of ensuring the integrity and quality of vector products, plasmids, and other critical raw materials. By leveraging the advantages of NGS over Sanger sequencing, manufacturers can ensure a higher standard of nucleic acid analysis, ultimately contributing to the reliability and efficacy of the final biopharmaceutical products.