Here, we employ the design of experiment (DoE) approach to analyze the influence and interdependence of factors (T7 RNA polymerase concentration, DNA template amount, and use of modified versus unmodified ribonucleotides) to develop an IVT performance model for assessing process performance using yield, integrity, and purity data of the mRNA pro...

Mar 18, 2022 · saRNAs are synthesized through in vitro transcription (IVT); however, this process has mainly been optimized for relatively short mRNAs. Methods: Here, we optimized the IVT process for long saRNAs, approximately 9.4 kb, through a design of experiment (DoE) approach to produce a maximal RNA yield and validated the optimal IVT method

Jan 8, 2021 · Here, we optimized the IVT process for long saRNAs, approximately 9.4 kb through a design of experiment (DoE) approach to produce a maximal RNA yield and validated the optimal IVT method on various sizes of RNA.

May 1, 2024 · Here we identified the optimal IVT conditions for short and long RNA transcripts through a DOE approach. We observed that saRNA requires lower temperature and Mg 2+ concentration to produce high quality transcripts and minimal dsRNA byproducts. mRNA quality, however, is not affected to the same extent by temperature and requires high Mg 2 ...

Testing the IVT reaction in HEPES and Tris buffer systems. Templates of 2.2 and 0.4 kB mRNA sizes (from plasmid and PCR DNA templates, respectively) were used for the mRNA synthesis.

In this study we determined optimal buffer conditions and nucleotide concentrations for maximizing the yield and quality of mRNA produced by in vitro transcription (IVT). By using design of experiments (DoE) principles and analytics, a variety of IVT reaction conditions were assessed for their impact on the IVT reactions.

This work will show the investigation of key mechanisms of mRNA in vitro transcription (IVT) as the first step in mRNA-based vaccine manufacturing on the proposed scalable machine, including the potential for accelerated candidate and process parameter optimization, to enable quick, scalable, risk-reduced, and resilient supply of critical ...

Jun 7, 2022 · We have developed a unique platform to obtain any specific size-range of LNPs from 60 to 180 nm satisfying pharmaceutical regulatory requirements for polydispersity index, sterility, dose...

Here, we employ the design of experiment (DoE) approach to analyze the influence and interdependence of factors (T7 RNA polymerase concentration, DNA template amount, and use of modified versus unmodified ribonucleotides) to develop an IVT performance model for assessing process performance using yield, integrity, and purity data of the mRNA ...

Here, we optimized the IVT process for long saRNAs, approximately 9.4 kb through a design of experiment (DoE) approach to produce a maximal RNA yield and validated the optimal IVT method on various sizes of RNA.