SLAS technologyOct 13, 2023

Using capillary electrophoresis to measure the size and purity of lab-made mRNA

mRNA Technology Weekly Brief ↗PubMed ↗DOI ↗

Updated Feb 19, 2026

Abstract

IVT mRNAs can range from less than 100 nt to longer than 9,000 nt.

Quality control is essential for producing IVT mRNA to ensure the correct length and absence of degradation.
Capillary gel electrophoresis offers high-resolution separation for assessing the purity of IVT mRNA.
The method allows differentiation between intact mRNA and truncated or degraded forms.
Specialized gels can analyze poly(A) tails, enabling the characterization of multiple Critical Quality Attributes.
Purity assessment methods can be applied to IVT mRNAs measuring either 6,000 or 9,000 nt.

Simplified

One result of the Covid-19 pandemic has been an increased awareness of IVT mRNA vaccines and the speed at which they can be produced for disease outbreaks. Currently the only approved IVT mRNA therapeutics are the Covid-19 vaccines, however IVT mRNA is being investigated for other non-Covid prophylactic vaccines, therapeutics, and therapeutic vaccines. IVT mRNAs can range from less than 100 nt in length to longer than 9,000 nt. When producing any IVT mRNA, quality control of the IVT mRNA is essential to ensure that the product is the correct length and does not contain truncated or degraded mRNA. Capillary gel electrophoresis provides high resolution separations of the IVT mRNA of interest from the degraded or truncated impurities allowing for the accurate purity assessment of IVT mRNA. Specialized capillary electrophoresis gels can also be used to provide analysis of purified poly(A) tails enabling characterization of multiple Critical Quality Attributes on a single platform. Here we describe methods for the purity assessment of IVT mRNAs through either 6,000 or 9,000 nt and determination of poly(A) tail length using different capillary gel electrophoresis methods.

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Competing interests

Declaration of Competing Interest Kyle Luttgeharm reports a relationship with Agilent Technologies Inc that includes: employment and equity or stocks. Whitney Pike reports a relationship with Agilent Technologies Inc that includes: employment and equity or stocks. Denise Warzak reports a relationship with Agilent Technologies Inc that includes: employment and equity or stocks.

PubMed

Using capillary electrophoresis to measure the size and purity of lab-made mRNA

Abstract

Full Text

Funding

Competing interests

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Abstract

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