GMP next generation sequencing identifies contaminants early and optimally delivers rapid, cost-effective results to assure product safety
Victor Muthu, PhD, Group Leader, BioPharma NextGen Sequencing; Marian L. McKee, PhD, Vice President, BioPharma Biosafety Testing, Eurofins BioPharma Product Testing
Next Generation Sequencing (NGS) technology has revolutionized the landscape of molecular biology, enabling high-throughput, accurate, and scalable sequencing for a variety of applications. At the forefront of this innovation, Eurofins BioPharma Product Testing (BPT) offers a comprehensive suite of NGS services, tailored to meet the needs of biopharmaceutical companies at all stages of product development. Our NGS offerings encompass sequence identity testing for plasmids, adeno-associated viruses (AAV), and lentivirus/retrovirus, along with adventitious viral detection services, ensuring quality control and safety in cell therapy, viral vector production, and related fields.
Identity tests are a crucial part of product testing packages submitted to regulatory agencies during the drug development and approval lifecycle. These tests include verifying the identity of plasmids and/or viral vectors that are critical raw materials, drug products (DP) or drug substances (DS) for cell and gene therapy manufacturing. Plasmid identity testing is particularly critical for confirming the presence of the correct insert and ensuring that the plasmid remains free of sequence variants or mutations. AAV and lentivirus/retrovirus sequence identity testing provides an analysis of packaged viral genomes, confirming that they are correctly assembled, free of unintended mutations, and maintain their intended therapeutic functionality. These tests are critical prior to releasing the plasmid or viral vector for downstream processes, including manufacturing.
Non-high throughput methods such as Sanger sequencing can be employed, but these are limited in the sequence coverage and depth that compromises the sensitivity of the assay. NGS is a high-throughput platform enabling rapid, cost-effective, parallel sequencing of DNA and RNA. Unlike Sanger sequencing, which analyzes one fragment at a time, NGS sequences millions of fragments simultaneously. NGS is the method of choice for identity testing as it provides the depth and sensitivity that are needed to identify the presence of any variants in a plasmid and/or viral vector nucleotide sequence. NGS also offers the ability for multiplex sequencing where samples can be batched and sequenced simultaneously, saving both costs and time, which is ideal for startups. Validated, GMP methods are ideal for release testing.
Eurofins BPT in Lancaster recently added GMP Sequence Identification Testing (SIT) by NGS to our testing portfolio. The method utilizes short-read sequencing performed on an Illumina® platform. The sequence obtained is compared to a client-provided reference sequence using bioinformatic analysis to identify any mismatches or variants, including insertions or deletions in the plasmid or viral vector sequence. The GMP method for Sequence Identity Testing has been validated in accordance with ICH Q2 (R2) as an identity method with additional attributes challenged to ensure the robustness of the method for detection of variants.
In addition to sequence identity testing, we also offer adventitious viral detection services (AAD). The presence of adventitious viruses in bioprocessing or gene therapy products can pose serious safety concerns and regulatory challenges. Our adventitious viral detection service leverages the sensitivity of NGS to identify a broad range of known and unknown viral contaminants that could potentially compromise product safety. This service is particularly valuable in the production of viral vectors, cell therapy, vaccines, and monoclonal antibody production, ensuring that products are free from contaminating adventitious viruses that could cause harm to patients or invalidate clinical trials. Our NGS-based adventitious viral detection services provide comprehensive evaluation of viral contamination, offering a higher level of detection sensitivity than traditional methods such as PCR or cell culture-based assays.
Our NGS services are powered by robust sequencing platforms, ensuring high throughput and unparalleled accuracy. We offer complete bioinformatics support, providing customers with detailed, actionable insights from the data analyses, including alignment and variant analysis for the SIT services, to contaminating virus identification for the AAD.
Eurofins’ NGS capabilities provide high quality, regulatory-accepted results to assure product safety and accelerate getting key materials into manufacturing and medicines to patients. Essential to a complete biosafety program, we enable clients to meet or exceed product milestones and to deliver critical therapies to patients faster than traditional methods.