Production of mRNA by in vitro transcription using a single-use system and ReadyToProcess™ WAVE™ 25 rocker Copy

Production of mRNA by in vitro transcription using a single-use system and ReadyToProcess™ WAVE™ 25 rocker

Single-use (SU) components were evaluated for production of messenger RNA (mRNA) by in vitro transcription (IVT). An initial test was performed using a Cellbag™ bioreactor container and a ReadyCircuit™ jumper Y manifold to investigate the presence of RNase activity and the suitability of the SU format for IVT reactions.

This test was followed by three separate mRNA transcriptions in Cellbag bioreactor containers at 150-mL scale using a linearized 7 kb DNA template that provides the sequence to be transcribed into 2000 nt mRNA. No difference in the assessed quality attributes was observed between IVT reactions performed in Cellbag bioreactor containers and control tubes. Yields were comparable and integrity analysis by capillary gel electrophoresis (CGE) showed no apparent sign of degradation.

These results indicate Cellbag bioreactor containers can be used for IVT reactions, at 150 mL scale but most likely also smaller and larger volumes. A single test of tangential flow filtration (TFF) was also performed using a Centramate™ omega cassette (100 kDa NMWCO). The recovery was > 90%. No sign of degradation was detected by CGE analysis and no loss of material in permeate was observed.

These results indicate that the chosen nominal molecular weight cutoff (NMWCO) is appropriate for this size of mRNA and larger constructs.

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Production of mRNA by in vitro transcription using a single-use system and ReadyToProcess™ WAVE™ 25 rocker

Thank You For Your Interest

Accurate Test Fixture Characterization and De-embedding