Sanger Sequencing Services
Sanger sequencing plays a vital role in verifying the identity of genetic materials utilized in biologics development and manufacturing. Widely recognized as the ‘Gold Standard,’ Sanger sequencing is extensively utilized to authenticate the DNA sequences of a wide array of biological raw materials, drug substances, and products.
Our Expertise
The Avance Biosciences team boasts over two decades of expertise in providing Sanger sequencing services. With a wealth of experience, we specialize in assisting clients with identification tests on a diverse range of genetic materials, including plasmids, viral vectors, mRNAs, PCR products, and beyond. Our Sanger sequencing services are offered as a platform method, validated to meet the requirements of GMP and GLP regulatory submissions.
Selected Applications:
Application
Description
Plasmid Identification
High-quality Sanger sequencing data is generated and assembled to create the full plasmid sequence. The consensus sequence is then compared to the reference sequence provided by the client, and any differences are reported.
mRNA DS Sequence Confirmation
Messenger RNA (mRNA) drug substances (DS) are sequenced following reverse transcription. The resulting consensus sequence is then compared to the reference sequence provided by the client, and any differences are reported.
Viral Vector Sequence Confirmation
As part of the viral vector release test, viral DNA or cDNA is sequenced to confirm the viral sequence, and the consensus sequence is compared to the reference sequence. Any variances or structural changes will be documented and reported.
Amplicon Genotyping
As the ‘Gold Standard’ for diagnostic assay cross-validation using clinical samples, the Sanger sequencing method is employed to verify mutations and indels in genes of interest. Before being utilized for clinical sample testing, both PCR primers and sequencing primers undergo thorough validation.
GMP QC Analysis Process
At Avance Biosciences, Sanger sequencing is validated for GMP identification tests. However, PCR and sequencing primers, critical components of these methods, are typically redesigned when testing samples from various therapeutic programs. We employ phase-appropriate processes to develop and validate identification methods based on Sanger sequencing. For GMP release of drug substances and drug products, we recommend the following procedures:
- Assay Development: PCR primers and sequencing primers will be designed and tested for assay specificity.
- Assay Validation: Assay specificity will be confirmed using positive and negative control samples.
- Sample Testing: Samples will be tested in compliance with GMP regulations using the validated methods.