A Comprehensive Lateral Flow Strip Assay for On‐Site mRNA Vaccine Quality Control in Decentralized Manufacturing

TL;DR

A portable and cost-effective lateral flow strip assay is developed for on-site quality control of mRNA vaccines, enabling rapid and accurate analysis in decentralized manufacturing settings.

Contribution

The development of a sequence-independent lateral flow strip assay for rapid and accurate on-site mRNA quality control in decentralized manufacturing.

Findings

The LFSA evaluates key mRNA quality attributes including 5' capping efficiency, integrity, and LNP encapsulation efficiency.

The assay provides real-time monitoring of mRNA stability under varying storage conditions and distinguishes between intact and degraded mRNA in LNPs.

Abstract

The rapid adoption of mRNA‐based vaccines highlights the critical need for on‐site quality control (QC) methods, particularly in low‐income countries with decentralized manufacturing. Existing techniques, such as liquid chromatography‐mass spectrometry (LC‐MS) and capillary electrophoresis (CE), are resource‐intensive, requiring specialized equipment and expertise. To address this, a comprehensive lateral flow strip assay (LFSA) has been developed to evaluate key mRNA quality attributes—5' capping efficiency, integrity, and lipid nanoparticles (LNPs) encapsulation efficiency. Leveraging antibody‐labeled fluorescence nanoparticles and polydeoxythymidine oligonucleotide to simultaneously probe both 5' cap and poly(A) tail, the LFSA offers sequence‐independent, rapid (15 min), and sensitive mRNA analysis. Validation against standard methods demonstrates comparable accuracy while significantly reducing sample requirements and operational complexity. Moreover, the LFSA enables real‐time monitoring of mRNA stability under varying storage conditions and provides precise encapsulation assessments, distinguishing between intact and degraded mRNA in LNPs. This portable and cost‐effective platform bridges critical gaps in mRNA QC, streamlines manufacturing workflows, and ensures mRNA therapeutic efficacy in diverse applications.

A rapid, portable lateral flow strip assay for on‐site mRNA quality control is developed, enabling sequence‐independent analysis of 5' capping, integrity, and LNPs encapsulation efficiency. This cost‐effective method offers real‐time stability monitoring, reduced sample requirements, and comparable accuracy to standard techniques, enhancing manufacturing workflows and ensuring mRNA therapeutic efficacy in decentralized settings.

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