Chromatographic separations for mRNA therapeutics: developing platform process for a new therapeutic modality

mRNA产品最近作为领先的COVID-19候选疫苗的突出地位对其制造提出了巨大挑战。成功将推动mRNA的使用进入一系列新的治疗应用和需要充分理解的制造processs.In体外转录（IVT）的DNA与噬菌体RNA聚合酶是目前最有效的方法来生产长序列特异性RNA。与基于细胞的合成相比，IVT是一种相对简单的程序，其允许模板定向合成任何序列的RNA分子，从短寡核苷酸到几种内切酶的寡核苷酸。IVT mRNA对于体内合成编码的治疗性蛋白质的重要性日益增加，需要制造纯mRNA产物。虽然该反应可以产生大量的RNA，但由于聚合酶的各种不需要的活性，它含有杂质。这些产品相关杂质包括脱帽mRNA、双链RNA和短单链RNA。他们显着影响产品质量，无论是通过阻碍mRNA的功效和最关注的激活免疫responses.This项目旨在解决这个纯化的挑战，首先了解产品相关杂质的性质在IVT反应，其次在此基础上的理解开发成本效益和可扩展的色谱分离，可以去除这些杂质的质量标准适合于临床应用。

The recent prominence of mRNA products as leading COVID-19 vaccine candidates places huge challenges on their manufacture. Success would drive the use of mRNA into a whole range of new therapeutic applications and the need for well understood manufacturing processes.In vitro transcription (IVT) of DNA with phage RNA polymerases is currently the most efficient method to produce long sequence-specific RNA. IVT is a relatively simple procedure when compared to cell based synthesis that allows for template-directed synthesis of RNA molecules of any sequence from short oligonucleotides to those of several kilobases. The increasing importance of IVT mRNA for synthesising the encoded therapeutic protein in vivo demands the manufacturing of pure mRNA products. While the reaction can yield large quantities of RNA, it contains impurities due to various unwanted activities of the polymerases. These product-related impurities include uncapped mRNA, double-stranded RNA, and short single stranded RNA. They significantly impact product quality, both by hampering mRNA efficacy and of most concern activating immune responses.This project seeks to address this purification challenge by first understanding the nature of product related impurities in IVT reactions and secondly on the basis of this understanding developing cost effective and scalable chromatographic separations that can remove these impurities to a specification suitable for clinical applications.