Field flow fractionation applications for the analysis of nanomaterials for health

Nanoparticle-based therapeutics represent a rapidly growing area of the pharmaceutical industry portfolio, with >55 nanomedicine-based products under clinical evaluation as of 2020, and a new wave of next-generation nanomedicines entering the clinical arena. While there are no universally accepted conventions for measuring nanomedicines, it is widely known that their safety and efficacy are directly correlated to their critical quality attributes, including particle size, drug loading, polydispersity, shape, as well as stability. Field-Flow Fractionation (FFF) as a high-resolution separation method has rapidly grown in interest over recent years, where it has been implemented for the separation and hyphenation with in-line detectors for the analysis of nanoparticle physicochemical parameters. Here, using a series of case studies we present the optimization of different FFF modes (frit inlet, asymmetric, and electrical field-flow fractionation), investigating various nanoscale materials
including polystyrene nanoparticles, immunoglobulin G (IgG), and lipid nanoparticles with variations in conditions based on the parameters. We investigated parameters influencing the effectiveness of FFF, including membrane composition, molecular-membrane weight cut-off (MWCO), channel
composition, carrier fluid and flow rate. We demonstrate that sample loss due to membrane interactions is a key challenge impacting the FFF-based separation of analytes. We present multiparametric experimental data showing the high-resolution analysis of nanoparticle and antibody formulation stability and compare them with orthogonal data, alongside the potential it holds for early stage analysis of bionanotherapeutics.