Many promising therapeutic agents have been produced because of the rapidly growing fields of cell and gene therapy. Although qPCR and ddPCR assays produce fast, reliable, and robust pharmacokinetic and pharmacodynamic profiles of these therapeutic agents, regulatory bodies have yet to create guidelines for the development and validation of qPCR and ddPCR methods, presenting a costly hurdle in the process of approving novel and life-saving treatments for clinical use. To address this, we adapted current regulatory guidelines for bioanalytical method validations (LBA) and referred to peer publications to develop a “Fit for purpose” GLP-compliant protocol for the development and validation of qPCR and ddPCR methods. In addition, to compare the efficacy of the two methods, we used shared matrix calibrators to cross-validate the results from the two methods. Our work provides a standardized and regulatory-friendly framework for qPCR, and further cements both qPCR and ddPCR as robust and reliable techniques.
Learning Objectives:
Upon completion, participant will be able to develop and validate methods for qPCR analysis and to cross-validate qPCR protocols with ddPCR.
Upon completion, participant will be able to optimize protocols for gDNA extraction to maximize DNA yields in a variety of tissues.
Upon completion, participant will be able to create an effective framework that ensures regulatory compliance for nascent scientific technologies that lack regulatory recommendations and standards.
