pSIM: A next generation in vitro mRNA potency assay.
- 1Technical Operation, CSL Seqirus, Holly Springs, NC 27540, USA.
- 2R&D, CSL Seqirus, 225 Wyman St, Waltham, MA 02451, USA.
Abstract
Potency testing for mRNA-lipid nanoparticle (LNP) vaccines remains challenging due to complex cell-based mechanisms and dependence on strain-specific antibodies. Here, we introduce a next-generation in vitro mRNA potency assay named as pSIM, pulsed stable isotope labeling by amino acids in cell culture (SILAC) internal dosing control (IDC) mass spectrometry (MS). The pSIM assay co-processes test and reference LNPs enabling effective concentration (EC50)-based or dosing-control-normalized linear slope readouts. We demonstrate its applicability across monocistronic and multivalent self-amplifying mRNA constructs for influenza and SARS-CoV-2, show an exploratory correlation between in vitro potency and in vivo neutralization in mice, and establish stability-indicating performance under heat stress, real-time storage, freeze-thaw, and RNA-lipid adduct formation. The assay is co-qualified for early phase quality control implementation to support phase-appropriate release and stability testing.
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BibTeX
@article{fox2026psim,
title = {{pSIM: A next generation in vitro mRNA potency assay.}},
author = {Marissa K Fox and Zhijun Cao and Hima Sapa and Zhichun Wang and William R Schutt and Raveen Rathnasinghe and Ying Zhang and Jiang Qian},
journal = {Molecular therapy. Advances},
year = {2026},
month = sep,
doi = {10.1016/j.omta.2026.201850},
url = {https://europepmc.org/article/MED/42908836},
note = {CC BY-NC-ND},
}
RIS
TY - JOUR
TI - pSIM: A next generation in vitro mRNA potency assay.
AU - Marissa K Fox
AU - Zhijun Cao
AU - Hima Sapa
AU - Zhichun Wang
AU - William R Schutt
AU - Raveen Rathnasinghe
AU - Ying Zhang
AU - Jiang Qian
PY - 2026
DA - 2026/09/21
JO - Molecular therapy. Advances
DO - 10.1016/j.omta.2026.201850
UR - https://europepmc.org/article/MED/42908836
AB - Potency testing for mRNA-lipid nanoparticle (LNP) vaccines remains challenging due to complex cell-based mechanisms and dependence on strain-specific antibodies. Here, we introduce a next-generation in vitro mRNA potency assay named as pSIM, pulsed stable isotope labeling by amino acids in cell culture (SILAC) internal dosing control (IDC) mass spectrometry (MS). The pSIM assay co-processes test and reference LNPs enabling effective concentration (EC50)-based or dosing-control-normalized linear slope readouts. We demonstrate its applicability across monocistronic and multivalent self-amplifying mRNA constructs for influenza and SARS-CoV-2, show an exploratory correlation between in vitro potency and in vivo neutralization in mice, and establish stability-indicating performance under heat stress, real-time storage, freeze-thaw, and RNA-lipid adduct formation. The assay is co-qualified for early phase quality control implementation to support phase-appropriate release and stability testing.
SN - 3117-387X
ER -