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Size-Exclusion Chromatography with Multi-angle Light Scattering for Elucidating Protein Aggregation Mechanisms

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Therapeutic Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 899))

Abstract

In this chapter, application of size exclusion chromatography with inline multi-angle light scattering (SEC-MALS) to protein systems is reviewed, in particular for its use in elucidating mechanistic details of net-irreversible aggregation processes. After motivating why SEC-MALS or analogous techniques are natural choices to interrogate such aggregating systems, the individual techniques (SEC and MALS) are reviewed briefly, as needed for the context of the remainder of the chapter. Illustrative examples are provided to highlight when and how SEC-MALS can be applied to test mass-action kinetic models for protein aggregation. Limitations of the technique, as well as recommendations for troubleshooting and potential errors in data interpretation are also provided.

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Author information

Authors and Affiliations

  1. Drug Product Science & Technology R&D, Bristol-Myers Squibb, New Brunswick, NJ, USA

    Erinc Sahin

  2. Department of Chemical Engineering, University of Delaware, Newark, DE, USA

    Christopher J. Roberts

Authors
  1. Erinc Sahin
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  2. Christopher J. Roberts
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Corresponding author

Correspondence to Christopher J. Roberts .

Editor information

Editors and Affiliations

  1. MedImmune, LLC, MedImmune Way One, Gaithersburg, 20878, USA

    Vladimir Voynov

  2. , Department of Physical Biochemistry, Biogen Idec, Cambridge Center 14, Cambridge, 02142, Massachusetts, USA

    Justin A. Caravella

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Sahin, E., Roberts, C.J. (2012). Size-Exclusion Chromatography with Multi-angle Light Scattering for Elucidating Protein Aggregation Mechanisms. In: Voynov, V., Caravella, J. (eds) Therapeutic Proteins. Methods in Molecular Biology, vol 899. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-921-1_25

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  • DOI: https://doi.org/10.1007/978-1-61779-921-1_25

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-920-4

  • Online ISBN: 978-1-61779-921-1

  • eBook Packages: Springer Protocols

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