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Publication Date

2018-05-14

First Advisor

Sarah J. Moore

Document Type

Honors Project

Degree Name

Bachelor of Science

Department

Engineering

Keywords

Cancer, Targeted therapy, Immunotherapy, Mesothelin, Protein engineering, Fibronectin, Cancer-Treatment, Cancer-Immunotherapy, Drug targeting, Mesothelium

Abstract

Current cancer treatment options can result in severe side effects for patients and do not prevent against cancer recurrence. Several targeted cancer therapies have been developed to reduce side effects but are not available for all cancer types. Immunotherapy, a subtype of targeted therapy, has been developed to direct the immune system to recognize a tumor, resulting in minimal side effects and lasting anti-tumor immunity for the patient. However, the immunosuppressive tumor microenvironment can make obtaining new anti-tumor immunity difficult. Therefore, the goal of this work is to develop a novel immunotherapy that harnesses pre-existing immunity to mount an attack against the tumor, obtaining anti-tumor immunity in the process. The proposed immunotherapy will consist of two components: a tumor-targeting agent and a viral epitope for which a patient has active immunity. This thesis aims to characterize conjugation methodologies between a tumor-targeting agent and a model viral epitope to assess feasibility of developing this novel therapeutic strategy. The tumor-targeting agent used in this work is the tenth domain of fibronectin type III (Fn3) that has been engineered to bind tumor cell surface biomarker, mesothelin (MSLN). The c-Myc peptide sequence was chosen to model a viral epitope. Three conjugation strategies were explored: direct protein fusion, C-terminal non-canonical amino acid incorporation for site specific conjugation, and C-terminal cysteine tag addition for polymer linker attachment. Both the C-terminal cysteine tag and the protein fusion Fn3 variants were successfully produced in microbial culture. Each variant maintained binding capability to MSLN. Additionally, when the direct fusion Fn3 variant was bound to MSLN on the surface of a tumor cell, an antibody was able to bind to the c-Myc tag, indicating feasibility of this novel therapeutic strategy.

Rights

2018 Samantha Rose Baierl. Access limited to the Smith College community and other researchers while on campus. Smith College community members also may access from off-campus using a Smith College log-in. Other off-campus researchers may request a copy through Interlibrary Loan for personal use.

Language

English

Comments

110 pages : illustrations (some color) Includes bibliographical references (pages 102-109)

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