Therapeutic proteins provide unique and critical treatments for many human diseases and conditions However, if a protein product cannot be stabilized adequately, its benefit to human health will never be realized. Proteins are highly susceptible to the formation of non-native aggregates and precipitates, which can cause adverse reactions in patients, ranging from immune response to anaphylactic shock and even death. Unfortunately, the mechanisms that control aggregation of therapeutic proteins are poorly understood. We have developed a tightly interwoven set of experiments, theories, and molecular level simulations to address our central hypothesis that the kinetics and thermodynamics of aggregation of therapeutic proteins are controlled by three main factors: conformational stability, colloidal stability, and interactions with interfaces. Each of these factors can be manipulated by changing solution conditions, protein structure, or both. To test our central hypothesis, we will manipulate protein conformational stability, protein-protein intermolecular interactions, and protein-surface interactions by employing innovative, state-of-the-art experimental methods. We have chosen a large set of model proteins in order to encompass a broad range of protein structural classes, molecular weights, and functionalities. These experiments, and their interpretation, will be complemented by innovative computer simulations and theoretical frameworks aimed at understanding the mechanisms that control protein aggregation at a molecular level. The Biotechnology Research Partnership team will be led by Prof. Ted Randolph, director of the Center for Pharmaceutical Biotechnology at the University of Colorado. Key scientific personnel on the team include Prof. Kristi Anseth (U. Colorado) an expert in biological applications of polymers, Prof. John Carpenter (U. Colorado Health Sciences Center), an expert in formulation of therapeutic proteins, Prof. Carol Hall, a leader in the field of computer simulations of protein aggregation, Prof. Kristi Kiick (U. Delaware), an expert in production of artificial proteins, Prof. Christopher Roberts (U. Delaware), an expert in modeling an analysis of protein aggregation, and Prof. Daniel Schwartz, a leader in studying protein behavior at interfaces. Relevance: To improve the safety of protein-based drugs, the problem of aggregation must be solved. The proposed research will discover how aggregation occurs, and develop strategies to prevent it. ? ? ?

National Institute of Health (NIH)
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Research Project (R01)
Project #
Application #
Study Section
Special Emphasis Panel (ZRG1-BCMB-G (91))
Program Officer
Henderson, Lori
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
University of Colorado at Boulder
Engineering (All Types)
Schools of Engineering
United States
Zip Code
Calderon, Christopher P; Daniels, Austin L; Randolph, Theodore W (2018) Deep Convolutional Neural Network Analysis of Flow Imaging Microscopy Data to Classify Subvisible Particles in Protein Formulations. J Pharm Sci 107:999-1008
Pardeshi, Neha N; Zhou, Chen; Randolph, Theodore W et al. (2018) Protein Nanoparticles Promote Microparticle Formation in Intravenous Immunoglobulin Solutions During Freeze-Thawing and Agitation Stresses. J Pharm Sci 107:1852-1857
Wang, Yiming; Gao, Yuan; Hill, Shannon E et al. (2018) Simulations and Experiments Delineate Amyloid Fibrilization by Peptides Derived from Glaucoma-Associated Myocilin. J Phys Chem B 122:5845-5850
Wang, Yiming; Hall, Carol K (2018) Seeding and cross-seeding fibrillation of N-terminal prion protein peptides PrP(120-144). Protein Sci 27:1304-1313
Daniels, Austin L; Randolph, Theodore W (2018) Flow Microscopy Imaging Is Sensitive to Characteristics of Subvisible Particles in Peginesatide Formulations Associated With Severe Adverse Reactions. J Pharm Sci 107:1313-1321
Calero-Rubio, Cesar; Ghosh, Ranendu; Saluja, Atul et al. (2018) Predicting Protein-Protein Interactions of Concentrated Antibody Solutions Using Dilute Solution Data and Coarse-Grained Molecular Models. J Pharm Sci 107:1269-1281
Sallam, Sahar; Dolog, Ivan; Paik, Bradford A et al. (2018) Sequence and Conformational Analysis of Peptide-Polymer Bioconjugates by Multidimensional Mass Spectrometry. Biomacromolecules 19:1498-1507
Haider, Michael J; Zhang, Huixi Violet; Sinha, Nairiti et al. (2018) Self-assembly and soluble aggregate behavior of computationally designed coiled-coil peptide bundles. Soft Matter 14:5488-5496
Gandhi, Aditya V; Pothecary, Mark R; Bain, David L et al. (2017) Some Lessons Learned From a Comparison Between Sedimentation Velocity Analytical Ultracentrifugation and Size Exclusion Chromatography to Characterize and Quantify Protein Aggregates. J Pharm Sci 106:2178-2186
Paik, Bradford A; Mane, Shivshankar R; Jia, Xinqiao et al. (2017) Responsive Hybrid (Poly)peptide-Polymer Conjugates. J Mater Chem B 5:8274-8288

Showing the most recent 10 out of 101 publications