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Determination of Protein Aggregation with Differential Mobility Analysis: Application to IgG Antibody

Published

Author(s)

Leonard F. Pease III, John T. Elliott, De-Hao D. Tsai, Michael R. Zachariah, Michael J. Tarlov

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 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 species are separated according to their electrophoretic mobility using a differential mobility analyzer. In this way, complete size distributions of protein species present, ranging from IgG monomers to pentamers, were obtained as ES-DMA spectra. The sizes of the IgG and IgG aggregates measured by DMA were found to correlate 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.
Citation
Biotechnology and Bioengineering
Volume
101
Issue
6

Keywords

Antibody aggregation, differential mobility analysis (DMA), electrospray (ES), irreversible aggregation, protein crystallography structure

Citation

Pease, L. , Elliott, J. , Tsai, D. , Zachariah, M. and Tarlov, M. (2008), Determination of Protein Aggregation with Differential Mobility Analysis: Application to IgG Antibody, Biotechnology and Bioengineering (Accessed April 15, 2024)
Created December 15, 2008, Updated February 19, 2017