Agilent Technologies, Ankeny, IA 50023, USA.
Agilent Technologies, Ankeny, IA 50023, USA.
SLAS Technol. 2023 Oct;28(5):369-374. doi: 10.1016/j.slast.2023.06.005. Epub 2023 Jul 2.
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 大流行的一个结果是人们越来越意识到 IVT mRNA 疫苗以及它们在疾病爆发时可以快速生产的速度。目前，唯一获得批准的 IVT mRNA 疗法是 Covid-19 疫苗，但是 IVT mRNA 正在被研究用于其他非 Covid 预防性疫苗、疗法和治疗性疫苗。IVT mRNAs 的长度可以从不到 100 个核苷酸到 9000 个核苷酸以上不等。在生产任何 IVT mRNA 时，都必须对 IVT mRNA 进行质量控制，以确保产品的长度正确，并且不含有截断或降解的 mRNA。毛细管凝胶电泳可从降解或截断的杂质中提供对感兴趣的 IVT mRNA 的高分辨率分离，从而可以准确评估 IVT mRNA 的纯度。专用的毛细管电泳凝胶也可用于分析纯化的 poly(A)尾，从而能够在单个平台上对多个关键质量属性进行表征。在这里，我们描述了通过 6000 或 9000 个核苷酸的 IVT mRNAs 的纯度评估方法，以及使用不同的毛细管凝胶电泳方法测定 poly(A)尾长度的方法。
