Publication Date

2017-5

Document Type

Honors Thesis

Degree Name

Bachelor of Science

Department

Engineering

Advisors

Sarah J. Moore

Keywords

Protein-drug conjugates, Mesothelin, Therapeutic targeting, Cancer, Protein engineering

Abstract

Chemotherapy has been the standard treatment for cancer for a while now. Targeted therapy has however been quickly gaining precedence in the cancer treatment landscape because of the increased efficacy and reduced toxicity associated with it. The main targeted therapy agents that have been approved for clinical use by FDA are antibody-based owing to the natural ability of antibodies to specifically bind and modify the activity of their respective antigens. To this end, antibody-drug conjugates (ADCs) have also been employed in treating cancer but a major limitation with such formulations is the fact that antibodies are large in size (~150 kDa) hence they can hardly penetrate through solid tumors.

To address this limitation, other protein-drug conjugates (PDCs), which can be over ten times smaller than ADCs are being considered as they have the potential to also achieve specific binding to a drug target. Currently there are no targeted therapeutics that are FDA approved for mesothelin-positive tumors like triple-negative breast, lung, ovarian and pancreatic cancers hence there is a need for such therapeutics. In this thesis, we seek to engineer a PDC that can be used to target mesothelin-expressing tumors. These efforts are based upon the modification of a

previously engineered targeting protein that specifically binds mesothelin. Primary amine groups on the protein and doxorubicin, a chemotherapeutic drug, are used in combination with an intermediate linker to successfully combine the protein and drug into a PDC.

High pressure liquid chromatography (HPLC) and SDS-PAGE are primarily employed to analyze the individual reactants in the conjugation reaction as well as the resultant product from the conjugation reaction. Further work needs to be done to accurately characterize the perceived protein-drug conjugate and analyze its potential utility as an anticancer agent for mesothelin expressing tumors.

Language

English

Comments

55 pages : illustrations (some color) Includes bibliographical references (pages 51-55)

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