Field-flow fractionation coupled to multi-angle light scattering (FFF-MALS) is a powerful analytical approach for the advanced characterization of nanomaterials (sizes < 1 μm). Contrary to size-exclusion chromatography, no stationary phase is required during the fractionation process, and the optimal separation range can be fine-tuned to measurement parameters. This flexibility enables reliable measurement of complex pharmaceutical products such as nanomedicines, monoclonal antibodies and vaccines. However, for quality control purposes the FFF-MALS protocol must satisfy regulatory needs and accepted standards. Here, we describe a validated FFF-MALS method in line with technical specification ISO/TS 21362 for the analysis of lipid-based nanoparticles (LNPs) encapsulating siRNA and mRNA.
LNPs for nucleic acid delivery, especially for short interfering RNA (siRNA) and messenger RNA (mRNA), have recently attracted extraordinary attention, and are expected to revolutionize the medical field. At the end of 2020 a milestone was reached with two vaccines against the ongoing COVID-19 pandemic, based on mRNA technology encapsulated in LNPs, approved by regulatory authorities in USA and Europe: BioNTech/Pfizer’s tozinameran and Moderna’s mRNA-1273. Apart from COVID-19 vaccines, further nucleic acid-based therapies are in development for a broad range of applications spanning immune-modulating agents, protein replacement therapies, regenerative medicine and gene-editing complexes, amongst others. Nanocarriers such as LNPs can protect active pharmaceutical ingredients, enhance bioavailability and thus improve safety and efficacy of novel therapies (1).
The use of lipid nanocarriers to deliver nucleic acid increases the complexity of the formulation and consequentially introduces the need for sophisticated analytics to ascertain a stable and safe drug product. Based on guidelines for drug products containing nanomaterials, including liposome characterization, the following parameters can be regarded as plausible critical quality attributes (CQAs): particle concentration, particle average size and polydispersity, nucleic acid loading levels, chemical stability and physical stability (aggregation propensity). Clearly, the advancement of robust analytical methods for CQA determination that are compliant with regulatory requirements is essential to streamline development and quality control.
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