Chapter 8: Biophysical Characterization and the Development of Therapeutic Proteins

  • Yangjie Wei
  • Nicholas R. Larson
  • Gang Hu
  • Prashant Kumar
  • C. Russell MiddaughEmail author
Part of the AAPS Advances in the Pharmaceutical Sciences Series book series (AAPS, volume 35)


During the formulation and development of therapeutic proteins, a thorough understanding of their structure and stability is essential. In this chapter, we discuss the most common methods used for this purpose and their individual advantages and disadvantages. This discussion is organized in terms of levels of structure (primary, secondary, tertiary, and quaternary including aggregation). Special attention is given to peptide mapping, circular dichroism, infrared and Fourier transform methods, ultraviolet absorbance, intrinsic and extrinsic fluorescence, and dynamic and static light scattering. Brief discussions concerning micro-flow imaging, nanoparticle tracking analysis, analytical ultracentrifugation, micro-calorimetry and hydrogen/deuterium exchange, and viscosity measurements are also included. Detailed examples are illustrated in figures and their legends.


Biophysical characterization Protein structure Protein stability Formulation Aggregation Circular dichroism Fluorescence Differential scanning calorimetry Light scattering Hydrogen/deuterium exchange 


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Copyright information

© American Association of Pharmaceutical Scientists 2020

Authors and Affiliations

  • Yangjie Wei
    • 1
    • 2
  • Nicholas R. Larson
    • 1
    • 2
  • Gang Hu
    • 1
    • 2
  • Prashant Kumar
    • 1
    • 2
  • C. Russell Middaugh
    • 1
    • 2
    Email author
  1. 1.Department of Pharmaceutical ChemistryMacromolecule and Vaccine Stabilization Center, University of KansasLawrenceUSA
  2. 2.Department of Pharmaceutical ChemistryUniversity of KansasLawrenceUSA

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