Optimization of In Vitro Transcription by Design of Experiment to Achieve High Self-Amplifying RNA Integrity.

ABSTRACT:

Background

Self-amplifying mRNA (saRNA) holds promising application prospects. However, due to the inclusion of a replicase sequence, its extended length leads to premature termination during in vitro transcription (IVT), resulting in poor product integrity. This study aims to optimize the IVT process for saRNA vaccines to enhance integrity, thereby addressing the key challenge in saRNA vaccine manufacturing.

Method

Guided by the Quality by Design (QbD) framework, Design of Experiment (DoE) methodology was employed to design diverse combinations of process parameters for IVT reactions. Predictive models were established to identify critical process parameters (CPPs) influencing integrity and yield. An optimized parameter set and process design space, meeting predefined yield

SUBMITTER: Hu C 

PROVIDER: S-EPMC12567852 | biostudies-literature | 2025 Oct

REPOSITORIES: biostudies-literature