Sequence Confirmation by NGS for Plasmid and Viral Vector Products
Confidence that the plasmid, viral vector, or RNA-based material behind your advanced cell and gene therapies is exactly what it’s supposed to be — from CMC development through GMP release. Results in under 2 weeks, from a validated, 21 CFR Part 11-ready pipeline built to handle the difficult cases — secondary/tertiary structure, high GC content, low-level impurities — that Sanger sequencing struggles with.
Built Around Your Therapeutic Program
Our services are organized around your therapeutic program and its regulatory milestones, from early CMC development through GMP release and beyond, so your NGS sequence confirmation sits alongside potency, qPCR/ddPCR, flow cytometry, immunoassay, and microbiology testing under one roof, run by a team that understands where your program is headed, not just what’s in the tube.
Reliable 21 CFR Part 11 Compliant NGS Analysis
Running NGS is easy, but meeting FDA 21 CFR Part 11 compliance is not. As the first company in the world to introduce Illumina NGS platform in GMP QC environments back in 2012, Avance Biosciences is a pioneer in applying NGS technology to support drug development and manufacturing. We deliver validated NGS data analysis pipelines with built-in audit trails to ensure data integrity and traceability. Our regulatory-ready approach supports high-quality, compliant results for biopharmaceutical development and manufacturing.
What We Test
Plasmid DNA and viral vector products (AAV, lentivirus, and other vector platforms)
Cell therapy and gene-edited cell products
mRNA and other RNA therapeutics
CRISPR/gene-editing constructs, including sgRNA
E. coli and other microbial production systems used in plasmid manufacturing
Improved NGS Sequence Confirmation with Avance as Your Testing Partner
How It Works
Every sample moves through a controlled, fully auditable process — with a QC checkpoint built into each stage, not just the final report.
Sequencing Difficult Templates Requires the Right Approach
Sanger sequencing often fails to produce reliable and accurate results for difficult-to-sequence templates. Any attempt to dissociate challenging secondary and tertiary structures ultimately destabilizes sequencing primer annealing, leading to failed Sanger sequencing results.
In contrast, random shearing in the NGS library preparation step can break up secondary structures, allowing successful sequencing through these regions, albeit at relatively lower coverage.
Although NGS ID is a platform method that does not require special sequencing primers or conditions for different samples, it is still necessary to perform a bridge study to define the difficult-to-sequence regions. This helps establish appropriate acceptance criteria for those regions.
While executing NGS for some of the identification tests is straightforward, ensuring compliance with FDA’s data integrity and Part 11 requirements during data analysis is challenging. Avance Biosciences has developed and validated numerous NGS data analysis pipelines, each with enforced audit trails to meet regulatory standards. This approach ensures the reliability and traceability of data generated, supporting rigorous quality assurance in biopharmaceutical development and manufacturing.
Technical Information
When a gene therapy company needed to verify plasmid identity, purity, and integrity for an IND submission, Avance Biosciences delivered a validated NGS-based solution. The assay enabled low-level variant detection and trace contaminant analysis, supporting FDA compliance and lot release.
