mRNA vaccine quality analysis using RNA sequencing

Helen M. Gunter, Senel Idrisoglu, Swati Singh, Dae Jong Han, Emily Ariens, Jonathan R. Peters, Ted Wong, Seth W. Cheetham, Jun Xu, Subash Kumar Rai, Robert Feldman, Andy Herbert, Esteban Marcellin, Romain Tropee, Trent Munro and Tim R. Mercer ()
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Helen M. Gunter: University of Queensland
Senel Idrisoglu: University of Queensland
Swati Singh: University of Queensland
Dae Jong Han: University of Queensland
Emily Ariens: University of Queensland
Jonathan R. Peters: University of Queensland
Ted Wong: Garvan Institute of Medical Research
Seth W. Cheetham: University of Queensland
Jun Xu: University of Queensland
Subash Kumar Rai: University of Queensland
Robert Feldman: COVID19 Vaccine Corporation Limited (CVC)
Andy Herbert: COVID19 Vaccine Corporation Limited (CVC)
Esteban Marcellin: University of Queensland
Romain Tropee: University of Queensland
Trent Munro: University of Queensland
Tim R. Mercer: University of Queensland

Nature Communications, 2023, vol. 14, issue 1, 1-12

Abstract: Abstract The success of mRNA vaccines has been realised, in part, by advances in manufacturing that enabled billions of doses to be produced at sufficient quality and safety. However, mRNA vaccines must be rigorously analysed to measure their integrity and detect contaminants that reduce their effectiveness and induce side-effects. Currently, mRNA vaccines and therapies are analysed using a range of time-consuming and costly methods. Here we describe a streamlined method to analyse mRNA vaccines and therapies using long-read nanopore sequencing. Compared to other industry-standard techniques, VAX-seq can comprehensively measure key mRNA vaccine quality attributes, including sequence, length, integrity, and purity. We also show how direct RNA sequencing can analyse mRNA chemistry, including the detection of nucleoside modifications. To support this approach, we provide supporting software to automatically report on mRNA and plasmid template quality and integrity. Given these advantages, we anticipate that RNA sequencing methods, such as VAX-seq, will become central to the development and manufacture of mRNA drugs.

Date: 2023
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DOI: 10.1038/s41467-023-41354-y

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