Characterization and solution structure of multi-domain proteins and protein complexes

Alina Vitaliyivna Kulakova*

*Corresponding author for this work

Research output: Book/ReportPh.D. thesis

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Abstract

Since the introduction of recombinant human insulin in 1982, protein-based therapeutics have become an essential part of medical treatment. Every year many protein-drugs are introduced into the market and numerous are under clinical trials. Despite all the advantages of protein-based pharmaceuticals, stabilization is very challenging. Under inappropriate conditions, they might lose their activity and induce severe adverse effects. Stabilization (formulation) of protein-drugs is one of the most critical, time-consuming and expensive steps in drug development. Many promising protein-drugs have failed clinical trials due to their stability challenges. Unfortunately, no general rules for the formulation process have been reported and it is not yet possible to predict the behavior of different proteins under different conditions.
Protein-excipient Interactions and Protein-Protein Interactions in formulation (PIPPI) is a consortium (EU grant nr 675074), which goal is to improve the molecular understanding behind protein stability and thereby of the formulation process. The strategy is to combine systematic investigations of the physicochemical behavior of different types of proteins with molecular interactions. The final goal is to combine all raw and analyzed data into a database that will become publicly available for the scientific community.
This Ph.D. thesis is a part of the PIPPI project, which focus is to investigate stability of multidomain proteins using small angle X-ray scattering. In this study, three different types of multidomain proteins were investigated: transferrin, albumin-neprilysin fusion protein, and monoclonal antibodies. All of them were systematically studied under different physicochemical conditions using high-throughput techniques. These studies were used to choose conditions for small angle X-ray scattering. The resulting data were combined with other measurements, such as MD simulation to provide a better understanding of stability on the molecular level. Despite the common trends in stability, all studied multidomain proteins show different behavior and molecular interactions. Additionally, this work shows that combination of multiple methods leads to a better understanding of protein stability.
Original languageEnglish
Place of PublicationKgs. Lyngby
PublisherTechnical University of Denmark
Number of pages155
Publication statusPublished - 2019

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