The work described in this dissertation encompasses
the design, production, purification, and testing of a set of novel
elastin-like polypeptides (ELP). Our aim in developing these proteins was for drug
delivery in Bladder Cancer (BC). BC has consistently shown the highest reoccurrence
rates of nearly any cancer, thus putting a heavy burden on patient quality of
life, and healthcare cost among its many other clinical challenges (Chapter 1).
To reduce such a burden, we sought to produce a targeted ELP which could be
used first as a contrast agent, and then provide the seminal work for a drug
delivery system. Throughout the development process, a unique organic
solvent-based extraction was developed for the expedited purification of ELP.
Beyond being orders of magnitude faster than any previously described method,
the organic extraction was able to remove key host contaminants (Chapter 2).
While great success was achieved using this facile organic extraction, complete
removal of organic solvents and minor contaminants became difficult and
laborious to achieve. These challenges post organic solvent extraction were
overcome by implementing a crucial precipitation step. The newly developed extraction-precipitation
method was then tested using various ELP to show the universality of this
purification method. (Chapter 3). Using our newly established method, we demonstrated
the clinical value by the purification of a targeting ligand, epidermal growth
factor (EGF), fused to an ELP. The performance of this construct was tested on
multiple bladder cancer models, as a potential contrast agent, and was compared
against non-targeted ELP (Chapter 4). A summation and forward look into
applications of my work, both the novel purification scheme, and the potential
impact of targeted ELP proteins in bladder cancer were explored (Chapter 5).