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April 4th, 2025
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InDevR, Inc.
molecular biology
biorxiv

Assay for Rapid Quantification of Capped and Tailed Intact mRNA

Gao, R. Y.Open in Google Scholar•Hu, T.Open in Google Scholar•Taylor, A. W.Open in Google Scholar•Lacey, R.Open in Google Scholar•Thomas, K. N.Open in Google Scholar•McCormick, C.Open in Google Scholar•Miller, D. F.Open in Google Scholar•Rowlen, K. L.Open in Google Scholar•Dawson, E. D.Open in Google Scholar

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, capping efficiency, and 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 cap and poly(A) tail. This 2-hour 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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