Date of Award

Spring 2018

Document Type

Thesis

Publisher

Santa Clara : Santa Clara University, 2018.

Department

Bioengineering

First Advisor

Zhiwen Zhang

Abstract

This Senior Design project seeks to manufacture an E. Coli based antibody for prostate cancer detection using unnatural amino acid incorporation. Current diagnostic techniques take advantage of the high binding specificity of monoclonal antibodies to detect the concentration of prostate specific antigen (PSA) in a blood sample. Production of this technology is lengthy and costly, while simultaneously incurring countless ethical problems related to animal welfare and accessibility. This project proposes a synthetic antibody that can be produced efficiently in single E. Coli cells and exhibits complementary binding to PSA. By incorporating an unnatural amino acid into a recombinant peptide sequence, binding affinity between substrate and antibody is substantially increased, effectively expanding the genetic code by introducing a novel function. Expression of the antibody is followed by extraction and purification of the synthetic antibody from cellular debris. Finally, the antibody’s binding strength and specificity to PSA is tested with respect to the monoclonal antibody with complementary binding to PSA. Assuming the synthetic antibody has greater or equivalent binding affinity than the currently used protein, this project will replace ethically questionable diagnostic techniques with a cheaper alternative.

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