ddPCR-Based Genomic Assay in Cell, Gene Therapies and Biomarker Research

Droplet digital PCR (ddPCR) is a widely used platform for detecting cell and gene therapy drugs, mutations and biomarkers because it provides absolute quantification of nucleic acids without the reference standard and calibration curve. The ddPCR-based genomic assay can be applied to the cellular kinetics for cell therapy, biodistribution for AAV-based gene therapy, vector genome titer measure, analytical development in the manufacturing process and translational biomarker research. In this panel, the presenters will discuss an overview of ddPCR-based genomic assays in cells, gene therapies and translational biomarker research and showcase unique case studies supporting drug discovery and biomarker analysis. The presenter will showcase the typical ddPCR assay is a 2-plex assay that detects a target chimeric antigen receptor (CAR) gene for monitoring the CAR cells and a reference gene for normalization in blood and biodistribution samples in tumor xenografted mice. It will also showcase the quantification of tumor-infiltrating regulatory T cell (Tregs) biomarkers involved in immune regulation and tolerance. The assay accurately quantifies the biomarker by one-step RT-ddPCR technology. The optimization process and challenges in dealing with the patient-derived samples, will be discussed. The assay demonstrated the linear dynamic range capable of detecting the low expression of the biomarker with minimal background noise in the cancer patient-derived samples. This ddPCR assay has significant potential for clinical research and therapeutic monitoring applications, particularly in cancer immunology and inflammatory diseases.