IVT Workflow Overview
In vitro transcription (IVT) is a method used to manufacture large quantities of mRNA. IVT is accomplished through either co-transcriptional or post-transcriptional capping (sometimes called enzymatic capping). In co-transcriptional capping, wild-type T7 (WT T7) RNA polymerase can achieve ~95% capping efficiency when optimized with trinucleotide cap analogs, and less than 80% capping efficiency with dinucleotide cap analogs (such as ARCA). Given the loss in capping efficiency, manufacturers prefer post-transcriptional capping with Vaccinia Capping Enzyme (VCE) when dinucleotide caps are used, as it can achieve >95% capping, but this also introduces additional process and purification steps.
Codexis enzymes are designed to address these trade-offs manufacturers face to improve overall process efficiency and quality. The Codex® HiCap RNA Polymerase is the first enzyme engineered to improve co-transcriptional capping efficiency using either trinucleotide or dinucleotide cap analogs, while achieving >95% capping efficiency with both cap analog formats.
Codex® HiCap RNA Polymerase
Improve mRNA capping efficiency
Produce capped synthetic mRNA at high yield and low immunogenicity with Codex® HiCap RNA Polymerase. Greater capping efficiency and the production of less double-stranded RNA byproduct deliver exceptional IVT results.
Read more about Codex® HiCap RNA PolymeraseScientific Highlights
An engineered T7 RNA polymerase for efficient co-transcriptional capping
Mathew Miller, Principal Scientist at Codexis, presented a poster at the 2022 Intl mRNA Health Summit highlighting how an engineered T7 RNA polymerase can overcome common challenges with capping efficiency and dsRNA impurities in IVT manufacturing of mRNA.
View poster