Senior Scientist Moderna Therapeutics Westborough, Massachusetts
RT-qPCR is used to quantitate circulating levels of mRNA from LNP encapsulated mRNA based therapeutics in serum to support clinical studies. There is no current guidance for application of RT-qPCR methods to measure mRNA from mRNA-LNP therapeutics including acceptance parameters for method validation and performance assessments during sample analysis.
We describe the development and qualification of RT-qPCR mRNA quantification from LNP-mRNA Therapeutics in human serum. We will describe 2 approaches
1. Extraction of mRNA from human serum spiked with mRNA-LNP drug product prior to RT-qPCR. 2. RT-qPCR measurement of mRNA from serum without extraction.
Measurement of mRNA without extraction generated high-quality data comparable to the extraction and has the advantage of not requiring an RNA extraction with less material input needed for the assay. The non-extraction method reduces sample preparation time, prevents sample loss or the need for back calculations, and provides a robust methodology for high throughput sample testing.
Learning Objectives:
Learn about mRNA-LNP therapeutics and RT-qPCR method development
Understand the method development process of RT-qPCR for PK studies in Clinical Develolpment
Can learn method development and qualification/validation parameters for the RT-qPCR method