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毛细管电泳法用于确定IVTmRNA尺寸和纯度的关键质量属性

Denise A Warzak¹, Whitney A Pike¹, Kyle D Luttgeharm¹

¹Agilent Technologies, Ankeny, IA 50023, USA.

SLAS technology

|October 13, 2023

概括

对于体外转录的信使RNA (IVT mRNA) 的质量控制至关重要. 毛细管凝电泳可以有效地评估IVTmRNA纯度和poly (A) 尾巴长度,用于治疗应用.

科学领域:

分子生物学分子生物学
生物技术是生物技术.
分析化学分析化学

背景情况:

随着COVID-19的流行,体外转录的信使RNA (IVT mRNA) 疫苗的重要性和快速生产能力得到了强调.
虽然目前仅限于COVID-19疫苗,IVT mRNA具有各种预防性疫苗,治疗药物和治疗性疫苗的潜力.
IVT mRNA分子的长度有很大的不同,从100个核酸以下 (nt) 到超过9,000nt.

研究的目的:

建立和描述用于评估IVTmRNA纯度的方法,特别是6000或9000nt长度的方法.
详细说明用于确定IVT mRNA的poly (A) 尾的长度的技术.
证明毛细管凝电泳 (CGE) 的实用性,用于表征IVT mRNA的关键质量属性.

主要方法:

利用毛细管凝电泳 (CGE) 来从切断或降解碎片等杂质中高分辨率地分离IVT mRNA.
采用专门的CGE凝来分析纯化的聚甲,从而实现了全面的表征.
开发并应用了针对纯度评估和多尾长度测定而定制的独特的CGE方法.

主要成果:

证明CGE通过有效地将全长IVTmRNA与降解产品分离,提供了准确的纯度评估.
展示了专门的CGE凝能够分析多尾巴长度的能力,这是一个关键的质量属性.
验证了CGE作为一个多功能平台,用于在单一系统上对IVTmRNA的多个关键质量属性进行表征.

结论:

毛细管凝电泳是确保IVTmRNA的质量和纯度的重要工具.
描述的CGE方法允许准确评估IVTmRNA长度和多A尾部完整性.
这种分析方法支持开发和质量控制各种基于IVT mRNA的治疗方法和疫苗.

关键词:

毛细管凝电泳发生在毛细管凝中.这种疫苗是Covid-19疫苗.在IVT的mRNA中.mRNA 关键质量属性 mRNA 关键质量属性的mRNA纯度.这是一种mRNA疗法.这是mRNA疫苗.

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