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Improved particle counting and size distribution determination of aggregated virus populations by asymmetric flow field-flow fractionation and multiangle light scattering techniques

Authors

  • Matt McEvoy,

    1. Dept. of Analytical Biochemistry, MedImmune, One MedImmune Way, Gaithersburg, MD 20878
    Current affiliation:
    1. 18162 Truffle Lane, Germantown, MD 20841
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  • Vladimir Razinkov,

    1. Dept. of Analytical Biochemistry, MedImmune, One MedImmune Way, Gaithersburg, MD 20878
    Current affiliation:
    1. Amgen, Inc., 1201 Amgen Court West, Seattle, WA 98119
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  • Ziping Wei,

    Corresponding author
    1. Dept. of Analytical Biochemistry, MedImmune, One MedImmune Way, Gaithersburg, MD 20878
    • Dept. of Analytical Biochemistry, MedImmune, One MedImmune Way, Gaithersburg, MD 20878
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  • Jose R. Casas-Finet,

    1. Dept. of Analytical Biochemistry, MedImmune, One MedImmune Way, Gaithersburg, MD 20878
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  • Guillermo I. Tous,

    1. Dept. of Analytical Biochemistry, MedImmune, One MedImmune Way, Gaithersburg, MD 20878
    Current affiliation:
    1. STIEFEL, a GSK company, 20 T.W. Alexander Drive, Research Triangle Park, NC 27709
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  • Mark A. Schenerman

    1. Dept. of Analytical Biochemistry, MedImmune, One MedImmune Way, Gaithersburg, MD 20878
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Abstract

A method using a combination of asymmetric flow field-flow fractionation (AFFFF) and multiangle light scattering (MALS) techniques has been shown to improve the estimation of virus particle counts and the amount of aggregated virus in laboratory samples. The method is based on the spherical particle counting approach given by Wyatt and Weida in 2004, with additional modifications. The new method was tested by analyzing polystyrene beads and adenovirus samples, both having a well-characterized particle size and concentration. Influenza virus samples were analyzed by the new AFFFF-MALS technique, and particle size and aggregate state were compared with results from atomic force microscopy analysis. The limitations and source of possible errors for the new AFFFF-MALS analysis are discussed. © 2010 American Institute of Chemical Engineers Biotechnol. Prog., 2010

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