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A single-nucleotide resolution capillary gel electrophoresis workflow for poly(A) tail characterization in the development of mRNA therapeutics and vaccines
Precise measurement method for mRNA tail length in developing mRNA treatments and vaccines
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Abstract
A workflow using capillary gel electrophoresis (CGE) for characterizing the poly(A) tail length of mRNA demonstrated resolution comparable to liquid chromatography coupled with mass spectrometry (LC-MS).
- The poly(A) tail length affects the stability and translation efficiency of mRNA, making it a critical quality attribute of mRNA therapeutics.
- Current methods like ion-pair reversed-phase liquid chromatography (IPRP-LC) can quantify mRNA with poly(A) tails but do not provide length information.
- High-resolution techniques such as LC-MS and next generation sequencing (NGS) can determine poly(A) tail length but are complex for routine release testing.
- The CGE method allows for quantification of each poly(A) length and provides relevant information for monitoring the quality of mRNA.
- This CGE method could serve as a suitable release testing approach for assessing poly(A) tail length in mRNA-based products.
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