High-Aspect Ratio Protein-Based Carriers for Delivery Applications
- Author(s): Bernard, Jenna Marie Lee
- Advisor(s): Francis, Matthew B.
- et al.
The work described in this dissertation aims to advance the early detection and therapeutic treatment of cancer. To do this, we seek to utilize protein-based carriers to deliver increased concentrations of cargo (imaging agents or drugs) to the tumor site that will allow for improved detection of early cancer states and/or a more focused delivery of therapeutics to the tumor.
Described herein is the production, characterization, and functionalization of two high-aspect ratio carriers: nanophage (Chapters 2-4) and a truncated IDP monomer (Chapter 5). To our knowledge, these structures have not yet been demonstrated for use in cancer delivery applications. In this work, a large emphasis was placed on the optimization of protein production and biochemical functionalization of these scaffolds for delivery applications.
The nanophage scaffold was explored for its genetic and chemical amenability to incorporate targeting moieties (scFv and cRGD, respectively) onto the phage coat proteins to create an “active targeting” agent that would facilitate cancer cell uptake (Chapter 4). Although the chemical incorporation of cRGD onto the nanophage was met with success, recent literature studies suggest that incorporation of targeting ligands onto a scaffold may not necessarily improve the nanocarrier accumulation in the tumor environment. Therefore, to obtain a better understanding of how the physical properties of a nanocarrier affect overall tumor accumulation, in vivo biodistribution studies were employed. Here, the proficiency of different carrier morphologies (sphere, disk, rod and disordered strand) to passively accumulate in the tumor environment was compared using glioblastoma tumor models. Moreover, in silico simulations and in vitro diffusion assays were used to further evaluate how the shape and size of these carriers might affect extravasation through a pore (Chapter 6).