One result of the Covid-19 pandemic has been an increased awareness of IVT mRNA vaccines and the speed at which they can be produced for disease outbreaks. Currently the only approved IVT mRNA therapeutics are the Covid-19 vaccines, however IVT mRNA is being investigated for other non-Covid prophylactic vaccines, therapeutics, and therapeutic vaccines. IVT mRNAs can range from less than 100 nt in length to longer than 9,000 nt. When producing any IVT mRNA, quality control of the IVT mRNA is essential to ensure that the product is the correct length and does not contain truncated or degraded mRNA. Capillary gel electrophoresis provides high resolution separations of the IVT mRNA of interest from the degraded or truncated impurities allowing for the accurate purity assessment of IVT mRNA. Specialized capillary electrophoresis gels can also be used to provide analysis of purified poly(A) tails enabling characterization of multiple Critical Quality Attributes on a single platform. Here we describe methods for the purity assessment of IVT mRNAs through either 6,000 or 9,000 nt and determination of poly(A) tail length using different capillary gel electrophoresis methods.

Covid-19 大流行的结果之一是提高了人们对 IVT mRNA 疫苗的认识以及针对疾病爆发生产疫苗的速度。目前唯一批准的 IVT mRNA 疗法是 Covid-19 疫苗，但正在研究 IVT mRNA 用于其他非 Covid 预防性疫苗、治疗剂和治疗性疫苗。IVT mRNA 的长度范围可以从小于 100 nt 到大于 9,000 nt。生产任何 IVT mRNA 时，IVT mRNA 的质量控制对于确保产品长度正确且不包含截短或降解的 mRNA 至关重要。毛细管凝胶电泳可将感兴趣的 IVT mRNA 与降解或截短的杂质进行高分辨率分离，从而可以准确评估 IVT mRNA 的纯度。专用毛细管电泳凝胶还可用于对纯化的聚 (A) 尾进行分析，从而能够在单一平台上表征多个关键质量属性。在这里，我们描述了通过 6,000 或 9,000 nt 评估 IVT mRNA 纯度的方法，以及使用不同毛细管凝胶电泳方法测定 Poly(A) 尾长的方法。