Design, Applications, and Processing of Synthetic Protein Nanoparticles

Nanomedicine- where a therapeutic is loaded into nanoparticles to increase therapeutic efficiency and improve patient outcomes- has long had the potential to revolutionize medicine. With all of their promise, nanoparticle carrier technologies have yet to make a significant clinical impact, emphasizing the need for new technologies and approaches. In this dissertation, electrohydrodynamic (EHD) co-jetting was used to develop various methods to create novel Synthetic Protein Nanoparticles (SPNPs), which were then applied to the delivery of therapeutic enzymes, and characterized using a microfluidic technique. It was found that SPNPs can be made from various proteins, such as Human Transferrin, Hemoglobin, and others, and that various macromers can be selected, such as a stimuli responsive NHS-Ester based macromer that can detect oxidative environments and show signs of degradation after being taken up by HeLa cells. SPNPs were then loaded with medically relevant enzymes, such as the antioxidant enzyme catalase. The enzymes showed high activity retention rates, with catalase SPNPs maintaining up to 82% of their original enzymatic activity. Additionally, antibody-targeted catalase SPNPs were able to protect REN cells in an inflammatory disease model. Next, an electrokinetic microfluidic system was adapted for the characterization of SPNPs based on their protein composition and anisotropy. The method was able to differentiate bicompartmental particles made from two different proteins from single compartment SPNPs made of an equivalent isotropic mixture of the same two proteins, with a significant voltage difference between the two particle types. Finally, preliminary work was conducted on using a small targeting molecule, meta-acetylenbenzylguanidine (MABG), for the treatment of neuroblastoma, and a system for validating MABG targeting in SK-N-BE(2) cells (a neuroblastoma cell line) was developed. Work done in this dissertation presents the development of multifunctional protein nanocarriers and lays the groundwork for the targeted delivery of active therapeutics using these particles.PhDBiomedical EngineeringUniversity of Michigan, Horace H. Rackham School of Graduate Studieshttp://deepblue.lib.umich.edu/bitstream/2027.42/163223/1/danqueve_1.pd