Article

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

  1. Leonard F. Pease III1,‡,
  2. Daniel I. Lipin2,
  3. De-Hao Tsai1,3,
  4. Michael R. Zachariah1,3,
  5. Linda H.L. Lua2,
  6. Michael J. Tarlov1,
  7. Anton P.J. Middelberg2,*

Article first published online: 18 AUG 2008

DOI: 10.1002/bit.22085

Issue

Biotechnology and Bioengineering

Biotechnology and Bioengineering

Volume 102, Issue 3, pages 845–855, 15 February 2009

Additional Information

How to Cite

Pease, L. F., Lipin, D. I., Tsai, D.-H., Zachariah, M. R., Lua, L. H., Tarlov, M. J. and Middelberg, A. P. (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: 845–855. doi: 10.1002/bit.22085

Author Information

  1. 1

    National Institute of Standards and Technology (NIST), Gaithersburg, Maryland

  2. 2

    Centre for Biomolecular Engineering, Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St Lucia, Brisbane 4072, Australia; telephone: +61-7-3346-4189; fax: +61-7-3346-4197

  3. 3

    The University of Maryland, College Park, Maryland

  1. ‡

    Leonard F. PeaseIII and Daniel I. Lipin contributed equally to this work.

*Centre for Biomolecular Engineering, Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St Lucia, Brisbane 4072, Australia; telephone: +61-7-3346-4189; fax: +61-7-3346-4197.

  1. †

    Reference to commercial equipment or supplies does not imply its endorsement by the US National Institute of Standards and Technology (NIST).

Publication History

  1. Issue published online: 17 DEC 2008
  2. Article first published online: 18 AUG 2008
  3. Accepted manuscript online: 18 AUG 2008 12:00AM EST
  4. Manuscript Accepted: 24 JUL 2008
  5. Manuscript Revised: 15 JUL 2008
  6. Manuscript Received: 12 MAR 2008

Funded by

  • Australian Research Council. Grant Numbers: FF0348465, DP0773111
  • Australian National Health and Medical Research Council. Grant Number: 409976

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Keywords:

  • virus;
  • influenza;
  • electrospray differential mobility analysis (ES-DMA);
  • asymmetric flow field-flow fractionation (AFFFF);
  • polyomavirus

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. Biotechnol. Bioeng. 2009; 102: 845–855. © 2008 Wiley Periodicals, Inc.

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