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High-Throughput Biophysical Analysis of Protein Therapeutics to Examine Interrelationships Between Aggregate Formation and Conformational Stability

  • Mini-Review
  • Theme: Aggregation and Interactions of Therapeutic Proteins
  • Published:
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Abstract

Stabilization and formulation of therapeutic proteins against physical instability, both structural alterations and aggregation, is particularly challenging not only due to each protein’s unique physicochemical characteristics but also their susceptibility to the surrounding milieu (pH, ionic strength, excipients, etc.) as well as various environmental stresses (temperature, agitation, lyophilization, etc.). The use of high-throughput techniques can significantly aid in the evaluation of stabilizing solution conditions by permitting a more rapid evaluation of a large matrix of possible combinations. In this mini-review, we discuss both key physical degradation pathways observed for protein-based drugs and the utility of various high-throughput biophysical techniques to aid in protein formulation development to minimize their occurrence. We then focus on four illustrative case studies with therapeutic protein candidates of varying sizes, shapes and physicochemical properties to explore different analytical challenges in monitoring protein physical instability. These include an IgG2 monoclonal antibody, an albumin-fusion protein, a recombinant pentameric plasma glycoprotein, and an antibody fragment (Fab). Future challenges and opportunities to improve and apply high-throughput approaches to protein formulation development are also discussed.

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  1. Rajoshi Chaudhuri

    Present address: Vaccine Production Program Laboratory, Vaccine Research Center/NIAID, National Institutes of Health, DHHS, 9 W. Watkins Mill Road, Suite 250, Gaithersburg, Maryland, 20878, USA

Authors and Affiliations

  1. Department of Pharmaceutical Chemistry, Macromolecule and Vaccine Stabilization Center, University of Kansas, Lawrence, Kansas, 66047, USA

    Rajoshi Chaudhuri, Yuan Cheng, C. Russell Middaugh & David B. Volkin

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  1. Rajoshi Chaudhuri
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Correspondence to David B. Volkin.

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Chaudhuri, R., Cheng, Y., Middaugh, C.R. et al. High-Throughput Biophysical Analysis of Protein Therapeutics to Examine Interrelationships Between Aggregate Formation and Conformational Stability. AAPS J 16, 48–64 (2014). https://doi.org/10.1208/s12248-013-9539-6

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