Successful Nucleic Acid Extraction: The Linchpin For PCR Bioanalysis

The AAPSE CHOP-TALK webinar, hosted by Eurofin’s clinical trial solutions and moderated by Timothy Lochmann, featured Dr. Mark Wissel, Director of R&D at URFIN’s Biopharma. Dr. Wissel discussed foundational strategies for nucleic acid extraction, a critical step for reliable PCR analysis in cell and gene therapy development. He compared leading extraction methods, including silica column- and bead-based approaches, emphasizing the importance of matching extraction technique to sample matrix—whether acellular like plasma and urine, or cellular like blood and PBMCs. Data from broad validation studies revealed that choice of extraction method, matrix type, and automation significantly influence assay sensitivity, consistency, and cost.

Dr. Wissel also addressed the role of internal controls and normalization to manage sample variability and improve recovery accuracy, citing industry references and a landmark PCR validation paper. He highlighted that high throughput, automated extraction is favored for minimizing human error and optimizing lab workflows. Case studies illustrated how extraction reagent quality and buffer pH impact PCR results, offering practical troubleshooting guidance. The webinar’s Q&A explored analyte-specific challenges, technology selection (QPCR vs. digital PCR), quantification techniques, and regulatory preferences for reporting results. Dr. Wissel recommended normalization using spiked controls for each sample, especially in studies with diverse matrices. He concluded that empirically-driven protocol design, thorough validation, and matrix-specific controls are key for successful PCR bioanalysis in clinical trials and gene therapy research.