Quality by design for mRNA platform purification based on continuous oligo-dT chromatography
Jixin Qu, Adithya Nair, George Muir, Kate A. Loveday, Zidi Yang, Ehsan Nourafkan, Emma N. Welbourne, Mabrouka Maamra, Mark J. Dickman, Zoltán Kis
Abstract
transcribed mRNA, which reduces losses, improves the efficiency of oligo-dT resin use, and intensifies the chromatography process. Furthermore, the quality by design (QbD) framework was used to establish a design space for the newly developed method. The optimization of process parameters (PPs), including salt type, salt concentration, load flow rate and mRNA load concentration both in batch and the continuous mode, achieved a greater than 90% yield (mRNA recovery) along with greater than 95% mRNA integrity and greater than 99% purity. The productivity of continuous chromatography was estimated to be 5.75-fold higher, and the operating cost was estimated 15% lower, when compared with batch chromatography. Moreover, the QbD framework was further used to map the relationship between critical quality attributes and key performance indicators as a function of critical process parameters and critical material attributes.