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

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.