Article

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

  1. Leonard F. Pease III1,
  2. John T. Elliott1,
  3. De-Hao Tsai1,
  4. Michael R. Zachariah1,2,*,
  5. Michael J. Tarlov1,*

Article first published online: 16 JUN 2008

DOI: 10.1002/bit.22017

Issue

Biotechnology and Bioengineering

Biotechnology and Bioengineering

Volume 101, Issue 6, pages 1214–1222, 15 December 2008

Additional Information

How to Cite

Pease, L. F., Elliott, J. T., Tsai, D.-H., Zachariah, M. R. and Tarlov, M. J. (2008), Determination of protein aggregation with differential mobility analysis: Application to IgG antibody. Biotechnology and Bioengineering, 101: 1214–1222. doi: 10.1002/bit.22017

Author Information

  1. 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. 2

    University of Maryland, College Park, Maryland 20742

*Correspondence: Michael R. Zachariah, Michael J. Tarlov, 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.).

  1. †

    Reference to commercial equipment, supplies, or software neither implies its endorsement by the National Institute of Standards and Technology (NIST) nor implies it to be necessarily best suited for this purpose.

Publication History

  1. Issue published online: 3 NOV 2008
  2. Article first published online: 16 JUN 2008
  3. Accepted manuscript online: 16 JUN 2008 12:00AM EST
  4. Manuscript Accepted: 16 MAY 2008
  5. Manuscript Revised: 15 MAY 2008
  6. Manuscript Received: 24 MAR 2008

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Keywords:

  • antibody aggregation;
  • protein crystallography structure;
  • differential mobility analysis (DMA);
  • electrospray (ES);
  • irreversible aggregation

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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