Enhancing mRNA-LNP Downstream Processing Via Single-Pass Tangential Flow Filtration

The emergence of lipid nanoparticle (LNP) encapsulated messenger ribonucleic acid (mRNA) vaccines has revolutionized genomic medicine offering significant advantages in delivery, safety, and scalability. A key step in LNP downstream processing is ultrafiltration and diafiltration via tangential flow filtration (TFF), which facilitates ethanol removal and product concentration. In this study, we investigated the feasibility of single-pass tangential flow filtration (SPTFF) as a continuous processing alternative for mRNA-LNPs. Comparative data were collected between conventional TFF and SPTFF, evaluating critical quality attributes including recovery, particle size, polydispersity, ethanol removal efficiency, and in vitro potency. Our findings demonstrate that SPTFF using 100 kDa regenerated cellulose cassettes is effective for concentration and ethanol removal while maintaining the LNP product quality attributes, when compared to conventional TFF. Notably, LNPs processed via SPTFF show a surprising 4.2-fold higher in vitro potency over LNPs that were produced by conventional TFF. This enhancement suggests that reduced dosing may be possible, potentially lowering toxicity risks associated with higher LNP quantities. Subsequent exploration of different SPTFF configurations and devices revealed performance variations across the design space.