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Biophysical Methods for Characterization of Antibody-Drug Conjugates

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Antibody-Drug Conjugates

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2078))

Abstract

Antibody-drug conjugates (ADC) are made up of three components: (1) a mAb specific to cells of choice, (2) a small molecule with desired end goal, and (3) a linker to covalently link drug molecule to the antibody. Bringing together the mAb, drug molecule, and the linker results in the formation of an immunoconjugate designed to selectively deliver the drug molecule to a cell of interest. Synergic effects of the mAb and drug molecule lead to destroying the target tumor cells while leaving the normal cells unharmed. However, the development of ADCs is associated with challenges due to the heterogeneity of the ADC molecules created from the conjugation process. Addition of the linker and drug moieties during processing as well as the hydrophobicity of the drug itself can lead to structural changes that may affect the stability and functional profile of the conjugated molecule. Furthermore, linkers site of attachment plays a major role in determining the conformational and colloidal properties of the ADCs. In this chapter, several characterization methods are introduced to determine the biophysical characteristics of the ADC. Protocols, data analysis as well as notes for circular dichroism, intrinsic fluorescence, ANS fluorescence, differential scanning calorimetry, and dynamic scanning fluorimetry are outlined in detail.

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Authors and Affiliations

  1. Pharmaceutical R&D, Pfizer, Inc., Chesterfield, MO, USA

    Vamsi Krishna Mudhivarthi & Jianxin Guo

Authors
  1. Vamsi Krishna Mudhivarthi
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  2. Jianxin Guo
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Corresponding author

Correspondence to Jianxin Guo .

Editor information

Editors and Affiliations

  1. Department of Pharmacy and Pharmaceutical Sciences, Binghamton University, Binghamton, NY, USA

    L. Nathan Tumey

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Mudhivarthi, V.K., Guo, J. (2020). Biophysical Methods for Characterization of Antibody-Drug Conjugates. In: Tumey, L. (eds) Antibody-Drug Conjugates. Methods in Molecular Biology, vol 2078. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9929-3_22

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  • DOI: https://doi.org/10.1007/978-1-4939-9929-3_22

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9928-6

  • Online ISBN: 978-1-4939-9929-3

  • eBook Packages: Springer Protocols

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