Assay for rapid quantification of capped and tailed intact mRNA

IF 4.5 3区医学 Q2 IMMUNOLOGY

Vaccine Pub Date : 2025-06-03 DOI:10.1016/j.vaccine.2025.127339

Rachel Y. Gao, Tianjing Hu , Amber W. Taylor , Randy Lacey, Keely N. Thomas, Caitlin McCormick , David F. Miller, Kathy L. Rowlen, Erica D. Dawson

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引用次数: 0

Abstract

The rapid development of mRNA vaccines since the COVID-19 pandemic has highlighted the potential of mRNA technology for vaccine and therapeutic applications. However, evaluating mRNA quality, including its integrity, 5′ capping efficiency, and 3′ poly(A) tailing, remains complex and challenging. Current analytical methods provide detailed insights, but are often limited by long analysis times, complex sample preparation, and the need for specialized expertise. The 5′ CapQ assay addresses these challenges and provides a novel, rapid, and straightforward method that offers a single, unique measurement of intact mRNA with both a 5'cap and 3'poly(A) tail. This 2-h benchtop-based assay provides quantitative or relative analysis, with high accuracy and precision without extensive sample processing. The 5′ CapQ assay offers a significant advancement in mRNA analytics to alleviate bottlenecks in applications including in vitro transcription (IVT) reaction optimization, post-transcriptional capping optimization, and assessment of batch-to-batch IVT bioprocess consistency.

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带帽和带尾完整mRNA的快速定量分析

自2019冠状病毒病大流行以来，mRNA疫苗的快速发展凸显了mRNA技术在疫苗和治疗应用方面的潜力。然而，评估mRNA的质量，包括其完整性、5 ‘封顶效率和3 ’聚(A)尾链，仍然是复杂和具有挑战性的。目前的分析方法提供了详细的见解，但往往受到长时间的分析，复杂的样品制备，以及需要专业知识的限制。5' CapQ检测解决了这些挑战，并提供了一种新颖、快速、直接的方法，可以对具有5'cap和3'poly(a) tail的完整mRNA进行单一、独特的测量。这种2小时的台式分析提供定量或相对分析，具有高精度和精密度，无需大量的样品处理。5 ' CapQ检测在mRNA分析方面取得了重大进展，缓解了包括体外转录（IVT）反应优化、转录后封顶优化和批间IVT生物过程一致性评估在内的应用瓶颈。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Vaccine 医学-免疫学

CiteScore

8.70

自引率

5.50%

发文量

992

审稿时长

131 days

期刊介绍： Vaccine is unique in publishing the highest quality science across all disciplines relevant to the field of vaccinology - all original article submissions across basic and clinical research, vaccine manufacturing, history, public policy, behavioral science and ethics, social sciences, safety, and many other related areas are welcomed. The submission categories as given in the Guide for Authors indicate where we receive the most papers. Papers outside these major areas are also welcome and authors are encouraged to contact us with specific questions.