title: Quantitative analysis of virus-like particle size and distribution by field-flow fractionation.
abstract: Asymmetric flow field-flow fractionation (AFFFF) coupled with multiple-angle light scattering (MALS) is a powerful technique showing potential for the analysis of pharmaceutically-relevant virus-like particles (VLPs). A lack of published methods, and concerns that membrane adsorption during sample fractionation may cause sample aggregation, have limited widespread acceptance. Here we report a reliable optimized method for VLP analysis using AFFFF-MALS, and benchmark it against dynamic light scattering (DLS) and transmission electron microscopy (TEM). By comparing chemically identical VLPs having very different quaternary structure, sourced from both bacteria and insect cells, we show that optimized AFFFF analysis does not cause significant aggregation, and that accurate size and distribution information can be obtained for heterogeneous samples in a way not possible with TEM and DLS. Optimized AFFFF thus provides a quantitative way to monitor batch consistency for new vaccine products, and rapidly provides unique information on the whole population of particles within a sample.
authors.affiliation: Centre for Biomolecular Engineering, Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St. Lucia, Brisbane 4072, Australia.
authors.forename: Yap P
authors.initials: YP
authors.lastname: Chuan
authors.forename: Yuan Y
authors.initials: YY
authors.lastname: Fan
authors.forename: Linda
authors.initials: L
authors.lastname: Lua
authors.forename: Anton P J
authors.initials: AP
authors.lastname: Middelberg
mesh_terms: D000465:Algorithms; D005591:Chemical Fractionation / Q000379:methods*; D044085:Microfluidics / Q000379:methods*; D009391:Nephelometry and Turbidimetry / Q000379:methods*; D010316:Particle Size; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014771:Virion / Q000302:isolation & purification* / Q000648:ultrastructure*