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...

2024年5月1日 · Messenger RNA (mRNA) and self-amplifying RNA (saRNA) vaccines against SARS-CoV-2 produced using in vitro transcription (IVT) were clinically approved in 2020 and 2022, respectively. While the industrial production of mRNA using IVT has been extensively optimized, the optimal conditions for saRNA have been explored to a lesser extent.

2021年1月8日 · 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. We found that magnesium has the highest impact on RNA yield with acetate ions enabling a higher yield than chloride ions.

本研究通过 实验设计 （DoE）方法优化了长saRNA（约9.4kb）的IVT过程，以产生最大RNA产量，并验证了不同大小RNA的最佳IVT条件。 主要缩略语 DoE：design of experiment，实验设计；

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.

借助经验证的实验设计（doe）和在转 染方面的专有技术，我们采用一种组合 方法来同时鉴定新型化合物并针对不同 rna有效负载进行剂型优化（图3）。 • 实现对潜在靶点的广泛筛选，以选择 最佳的脂质纳米颗粒(lnp) • 通过doe优化lnp制剂工艺参数 合成 新脂质

To test if the choice of buffering system is a critical factor for the overall performance of the IVT reaction, we conducted an experiment with HEPES and Tris buffer. Both Tris and HEPES buffers resulted in comparably high mRNA yield, independent of

2024年3月19日 · 采用 DOE，探索合成 mRNA 与 saRNA 的最佳反应条件. 采用确定性筛选设计 （ Definitive Screening Design） 评估对 mRNA 与 saRNA 的质量产量造成关键差异的 IVT 参数。研究者对每个 RNA 分子进行了 25 次 IVT 反应，重复三次。这样做会显着减少运行的 IVT 反应数量。

saRNA合成生产同样通过典型的 体外转录 反应（IVT）合成： RNA酶抑制剂（RNase Inhibitors），抑制IVT反应体系里可能存在的RNase。 无机焦磷酸酶（Pyrophosphatase），降解IVT反应积累的焦磷酸。 镁离子（Magnesium），T7 RNA聚合酶的辅因子。 PH缓冲buffer，含最佳浓度的抗氧化剂和多胺。 当前IVT反应体系基本上是从适用于合成100nt的IVT合成体系优化发展而来，并非专门替合成saRNA量身制定。 saRNA长度高达9.4kb，含有更多的二级结构，用来 …

2022年6月7日 · We applied design of experiment (DoE) methodology and identified the critical process parameters to establish the process for global application. Cross-point validation within the response map of...