Quantitative characterization of virus-like particles by asymmetrical flow field flow fractionation, electrospray differential mobility analysis, and transmission electron microscopy

Pease, Leonard F., Lipin, Daniel I., Tsai, De-Hao, Zachariah, Michael R., Lua, Linda H. L., Tarlov, Michael J. and Middelberg, Anton P. J. (2009) Quantitative characterization of virus-like particles by asymmetrical flow field flow fractionation, electrospray differential mobility analysis, and transmission electron microscopy. Biotechnology and Bioengineering, 102 3: 845-855. doi:10.1002/bit.22085


Author Pease, Leonard F.
Lipin, Daniel I.
Tsai, De-Hao
Zachariah, Michael R.
Lua, Linda H. L.
Tarlov, Michael J.
Middelberg, Anton P. J.
Title Quantitative characterization of virus-like particles by asymmetrical flow field flow fractionation, electrospray differential mobility analysis, and transmission electron microscopy
Journal name Biotechnology and Bioengineering   Check publisher's open access policy
ISSN 0006-3592
1097-0290
Publication date 2009-02-15
Year available 2008
Sub-type Article (original research)
DOI 10.1002/bit.22085
Volume 102
Issue 3
Start page 845
End page 855
Total pages 11
Editor D. S. Clark
Place of publication Hoboken, New Jersey, United States of America
Publisher John Wiley and Sons
Language eng
Subject 100302 Bioprocessing, Bioproduction and Bioproducts
Abstract Here we characterize virus-like particles (VLPs) by three very distinct, orthogonal, and quantitative techniques: electrospray differential mobility analysis (ES-DMA), asymmetric flow field-flow fractionation with multi-angle light scattering detection (AFFFF-MALS) and transmission electron microscopy (TEM). VLPs are biomolecular particles assembled from viral proteins with applications ranging from synthetic vaccines to vectors for delivery of gene and drug therapies. VLPs may have polydispersed, multimodal size distributions, where the size distribution can be altered by subtle changes in the production process. These three techniques detect subtle size differences in VLPs derived from the non-enveloped murine polyomavirus (MPV) following:(i) functionalization of the surface of VLPs with an influenza viral peptide fragment; (ii) packaging of foreign protein internally within the VLPs; and (iii) packaging of genomic DNA internally within the VLPs. These results demonstrate that ES-DMA and AFFFF-MALS are able to quantitatively determine VLP size distributions with greater rapidity and statistical significance than TEM, providing useful technologies for product development and process analytics.
Keyword Virus
Influenza
Electrospray differential mobility analysis (ES-DMA)
Asymmetric flow field-flow fractionation (AFFFF)
Polyomavirus
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ
Additional Notes Published online 18 August 2008 in Wiley InterScience

Document type: Journal Article
Sub-type: Article (original research)
Collection: Australian Institute for Bioengineering and Nanotechnology Publications
 
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Created: Thu, 03 Sep 2009, 08:57:04 EST by Mr Andrew Martlew on behalf of Aust Institute for Bioengineering & Nanotechnology