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Quantitative analysis of virus‐like particle size and distribution by field‐flow fractionation
Author(s) -
Chuan Yap P.,
Fan Yuan Y.,
Lua Linda,
Middelberg Anton P.J.
Publication year - 2007
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.21710
Subject(s) - field flow fractionation , dynamic light scattering , fractionation , light scattering , biological system , multiangle light scattering , population , particle (ecology) , chemistry , chromatography , scattering , materials science , nanotechnology , analytical chemistry (journal) , nanoparticle , optics , biology , physics , ecology , demography , sociology
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. Biotechnol. Bioeng. 2008;99: 1425–1433. © 2007 Wiley Periodicals, Inc.