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Recombinant proteins and antibodies have both diagnostic and therapeutic uses with biological therapeutics being one of the largest markets in the pharmaceutical industry. The work in this thesis details a series of studies examining novel applications of antibodies and recombinant proteins in the treatment of cancer. Initially, two novel fusion tags were evaluated for their ability to overcome some of the problems associated with the expression of recombinant proteins in Escherichia coli to use as immunogens to raise antibodies. Both of the fusion tags where shown to be effective as N-terminal fusion tags for the expression and purification of target proteins and may prove useful in circumventing expression and purification issues associated with E. coli as an expression host. 1)1 the second part of the project, amphiregulin (AREG) and heparin binding epidermal growth factor (HBEGF) recombinant proteins were expressed in E. coli and used to produce monoe/onal antibodies. The AREG antibody and HBEGF/AREG cross-specific antibodies showed anti-proliferative effects that were enhanced by chemotherapy with significant effects on cell invasion also observed. The antibodies enabled the validation of targeting AREG and HBEGF as a potential cancer therapy. Finally, a new fusion protein therapeutic composed of the two cathepsin peptides was developed and examined. Inhibitory effects were observed on the enzymatic activity of both cathespins Band S as well as a reduction in cell invasion. This inhibitory molecule may have utility in the treatment of cancer given the role played by both proteases in tumourigenesis. In this study, the use of recombinant proteins as immunogens for generation of monoe/onal antibodies as well as a targeted therapy in their own right has been demonstrated. Both inhibitory molecules developed in this study have potential to be successful biological therapies and will be examined further. |
