Real-Time PCR Services
Real-time quantitative PCR plays a pivotal role in drug development and manufacturing processes, providing valuable insights into gene expression, biomarkers, pharmacogenomics, quality control, process monitoring, and viral vector production. Its accuracy, sensitivity, and versatility make it an indispensable tool for advancing new therapeutics and ensuring the safety and efficacy of pharmaceutical products.
Our Expertise
The Avance Biosciences team boasts over two decades of expertise in providing real-time PCR services. Our scientists are proficient in TaqManTM assay design and have extensive experience validating various assays in compliance with FDA and ICH guidelines for analytical and bioanalytical method validation. With a proven track record, our team excels in providing creative solutions to address our clients’ specific needs, ensuring efficient, sensitive, and reproducible assays.
Selected Applications
Application
Description
CHO Copy Number Analysis
Two TaqMan™ assays are designed and validated to determine the copy number of integrated genes, normalized to an endogenous gene. This analysis reports the copies of genes integrated in Master Cell Banks (MCB), Working Cell Banks (WCB), and End-of-Production Cells (EOPC) for genetic stability analysis.
CAR-T CNV Analysis
Two TaqMan™ assays are designed and validated to determine the copy number of lentivirus-inserted or retrovirus-inserted therapeutic genes, normalized to an endogenous gene. This analysis is conducted for CAR-T cell release testing, with a one-week turnaround time.
RCL Testing
A sensitive TaqMan™ assay is designed and validated to determine the level of the VSVG plasmid fragment in CAR-T cells, assessing the risk of replication-competent lentivirus.
CGT Biodistribution
We will design and validate sensitive TaqMan™ assays to determine the distribution and persistence of viral vectors (RNA or DNA) or therapeutic cells in various animal models. These assays undergo validation according to FDA requirements for bioanalytical method validation, including validation of extraction recovery.
Gene Expression Analysis
RNA expression levels after drug treatment in both animal or human subjects are analyzed using a target gene-specific assay and an endogenous normalizing assay. Depending on the analysis requirements, both the standard curve method and the ΔΔCt method can be utilized.
TaqMan Genotyping
As the ‘Gold Standard’ for diagnostic assay cross-validation using clinical samples, TaqMan™ assays are employed to verify mutations in genes of interest. Before being utilized for clinical sample testing, assays undergo validation according to FDA requirements for bioanalytical methods.
Regulated Analysis Process
At Avance Biosciences, Real-Time PCR assays are validated either as a platform method or designed and validated as custom assays for GMP/GLP testing. In cases of platform methods, a phase-appropriate partial validation is typically recommended before sample testing. Custom assays, including those for GMP release testing and for animal or human studies, undergo independent validations in accordance with ICH and FDA requirements on analytical and bioanalytical method validation.
- Assay Development: TaqMan™ primers and probes are designed and tested for specificity and PCR efficiency. Additionally, the linearity, dynamic range, limit of quantification (LOQ), and limit of detection (LOD) will be assessed.
- Assay Validation: Assay validation will adhere to a pre-approved validation plan, ensuring compliance with the requirements outlined in ICH guidelines and FDA guidance documents for analytical and bioanalytical method validation.
- Sample Testing: Samples are handled with utmost care to prevent the risk of cross-contamination. Separate laboratory rooms, including an extraction room, PCR master mix room, PCR template room, and post-PCR room, are designated for each stage of sample analysis.