Abstract
The neonatal Fc receptor (FcRn) plays a key role in determining the pharmacokinetic behavior of therapeutic monoclonal antibodies (mAbs). FcRn-mediated intracellular trafficking mechanisms extend the half-lives of mAbs by rescuing them from lysosomal degradation and contribute to their transportation from the vascular space to tissue compartments such as placenta and mucosal surfaces. It is important to characterize the FcRn interactions of therapeutic mAbs and Fc-fusion proteins due to its potential impact on their in vivo pharmacokinetic properties such as clearance and half-life. In this chapter, we describe protocols for two cell-based assays that measure the total function of FcRn which involves pH-dependent association and dissociation with IgG-Fc, as well as FcRn-mediated intracellular trafficking parameters. These assays are suitable for characterization of FcRn interactions with therapeutic mAbs and Fc-fusion proteins for the purpose of assessing lot-to-lot consistency and the structural and functional integrity of the Fc domain. In addition, they may serve as cost-effective screening tools for the evaluation of mAb-based drug candidates during lead selection and optimization for desired pharmacokinetic properties.
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Acknowledgments
The authors would like to thank Patricia Y. Siguenza for her support of the project, Dr. Lynn Kamen for providing valuable comments on the manuscript, and colleagues at the Core Biophysical Characterization and Reagent Facility, and the Critical Reagents System for providing critical reagents.
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Chung, S., Lin, Y.L., Nguyen, V., Liu, C. (2022). Methods for Functional Characterization of FcRn Interactions with Therapeutic Antibodies and Fc-Fusion Proteins. In: Houen, G. (eds) Therapeutic Antibodies. Methods in Molecular Biology, vol 2313. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1450-1_18
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DOI: https://doi.org/10.1007/978-1-0716-1450-1_18
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Publisher Name: Humana, New York, NY
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Online ISBN: 978-1-0716-1450-1
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