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Current and Emerging Technologies to Characterize the Higher-Order Structure of Monoclonal Antibodies



John Marino, Robert G. Brinson, Jane E. Ladner, David Travis Gallagher, Luke Arbogast, Richard Huang, Jeffrey W. Hudgens, Elyssia S. Gallagher


In contrast to small molecule therapeutics whose conformations can be absolutely defined by constitution and stereochemistry, biopharmaceuticals are distinguished by the requirement for folding into higher order structures (secondary, tertiary, and quaternary) for therapeutic function. While proper folding of a protein biologic is critical for drug efficacy, misfolding may impact drug safety by eliciting unwanted immune or other off-target patient responses. In this chapter, we review current and emerging technologies for high-resolution characterization of the structure and dynamics of monoclonal antibodies (mAbs) with a focus on techniques that can provide data at and or near atomic resolution, such as X-ray crystallography, nuclear magnetic resonance (NMR) and HDX-mass spectrometry. Application of these techniques is illustrated using the NIST mAb.
Current, State of the Art, and Emerging Technolgies for the Analysis of Monoclonal Antibodies
Publisher Info
American Chemical Society, Washington, DC


mAb, NMR, X-ray Crystallography, HDX-MS, higher order structure


Marino, J. , Brinson, R. , Ladner, J. , Gallagher, D. , Arbogast, L. , Huang, R. , Hudgens, J. and Gallagher, E. (2015), Current and Emerging Technologies to Characterize the Higher-Order Structure of Monoclonal Antibodies, American Chemical Society, Washington, DC, [online], (Accessed June 24, 2024)


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Created October 14, 2015, Updated October 12, 2021