Determination of protein aggregation with differential mobility analysis: Application to IgG antibody

Authors

  • Leonard F. Pease III,

    1. National Institute of Standards and Technology (NIST), 100 Bureau Drive MS 8362, Gaithersburg, Maryland 20899; telephone: 301-975-2058 (M.J.T.)/301-405-4311 (M.R.Z.); fax: 301-975-2643 (M.J.T./M.R.Z.)
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  • John T. Elliott,

    1. National Institute of Standards and Technology (NIST), 100 Bureau Drive MS 8362, Gaithersburg, Maryland 20899; telephone: 301-975-2058 (M.J.T.)/301-405-4311 (M.R.Z.); fax: 301-975-2643 (M.J.T./M.R.Z.)
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  • De-Hao Tsai,

    1. National Institute of Standards and Technology (NIST), 100 Bureau Drive MS 8362, Gaithersburg, Maryland 20899; telephone: 301-975-2058 (M.J.T.)/301-405-4311 (M.R.Z.); fax: 301-975-2643 (M.J.T./M.R.Z.)
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  • Michael R. Zachariah,

    Corresponding author
    1. National Institute of Standards and Technology (NIST), 100 Bureau Drive MS 8362, Gaithersburg, Maryland 20899; telephone: 301-975-2058 (M.J.T.)/301-405-4311 (M.R.Z.); fax: 301-975-2643 (M.J.T./M.R.Z.)
    2. University of Maryland, College Park, Maryland 20742
    • National Institute of Standards and Technology (NIST), 100 Bureau Drive MS 8362, Gaithersburg, Maryland 20899; telephone: 301-975-2058 (M.J.T.)/301-405-4311 (M.R.Z.); fax: 301-975-2643 (M.J.T./M.R.Z.).
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  • Michael J. Tarlov

    Corresponding author
    1. National Institute of Standards and Technology (NIST), 100 Bureau Drive MS 8362, Gaithersburg, Maryland 20899; telephone: 301-975-2058 (M.J.T.)/301-405-4311 (M.R.Z.); fax: 301-975-2643 (M.J.T./M.R.Z.)
    • National Institute of Standards and Technology (NIST), 100 Bureau Drive MS 8362, Gaithersburg, Maryland 20899; telephone: 301-975-2058 (M.J.T.)/301-405-4311 (M.R.Z.); fax: 301-975-2643 (M.J.T./M.R.Z.).
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Abstract

Here we describe the use of electrospray differential mobility analysis (ES-DMA), also known as gas-phase electrophoretic mobility molecular analysis (GEMMA), as a method for measuring low-order soluble aggregates of proteins in solution. We demonstrate proof of concept with IgG antibodies. In ES-DMA, aqueous solutions of the antibody protein are electrosprayed and the various aerosolized species are separated according to their electrophoretic mobility using a differential mobility analyzer. In this way, complete size distributions of protein species present from 3 to 250 nm can be obtained with the current set up, including distinct peaks for IgG monomers to pentamers. The sizes of the IgG and IgG aggregates measured by DMA were found to be in good agreement with those calculated from simple models, which take the structural dimensions of IgG from protein crystallographic data. The dependence of IgG aggregation on the solution concentration and ionic strength was also examined, and the portion of aggregates containing chemically crosslinked antibodies was quantified. These results indicate that ES-DMA holds potential as a measurement tool to study protein aggregation phenomena such as those associated with antibody reagent manufacturing and protein therapeutics. Biotechnol. Bioeng. 2008;101: 1214–1222. © 2008 Wiley Periodicals, Inc.

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