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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 by Deanna Di Grandi; Daniel M. Dayeh; Keerat Kaur; Yizhuo Chen; Steven Henderson; Youmi Moon; Arijit Bhowmick; Peter M. Ihnat; Yue Fu; Kathir Muthusamy; Nisha Palackal; Erica A. Pyles is a Chemistry article available to read on EtoBox.

What is A single-nucleotide resolution capillary gel electrophoresis workflow for poly(A) tail characterization in the development of mRNA therapeutics and vaccines about?

The 3′ poly(A) tail is an important component of messenger RNA (mRNA). The length of the poly(A) tail has direct impact on the stability and translation efficiency of the mRNA molecule and is therefore considered to be a critical quality attribute (CQA) of mRNA-based therapeutics and vaccines. Various analytical methods have been developed to monitor this CQA. Methods like ion-pair reversed-phase liquid chromatography (IPRP-LC) can be used to quantify the percentage of mRNA with poly(A) tail but fail to provide further information on the actual length of poly(A). High-resolution methods such as liquid chromatography coupled with mass spectrometry (LC-MS) or next generation sequencing (NGS) can separate poly(A) tail length by one nucleotide (n/n + 1 resolution) but are complicated to implement for release testing of manufactured mRNA. In this study, a workflow utilizing capillary gel electrophoresis (CGE) for characterizing the poly(A) tail length of mRNA was developed. The CGE method demonstrated resolution comparable with the LC-MS method. With UV detection and the addition of poly(A) length markers, this method can provide poly(A) tail length information and can also provide quan

Who reads A single-nucleotide resolution capillary gel electrophoresis workflow for poly(A) tail characterization in the development of mRNA therapeutics and vaccines?

Author

Deanna Di Grandi; Daniel M. Dayeh; Keerat Kaur; Yizhuo Chen; Steven Henderson; Youmi Moon; Arijit Bhowmick; Peter M. Ihnat; Yue Fu; Kathir Muthusamy; Nisha Palackal; Erica A. Pyles

Publisher

Elsevier BV

Published

2023

Language

EN

Field

Chemistry (Physical Sciences)