Development and applications of novel approaches for monitoring antibody generation to prostate cancer-related antigens
Lemass, Darragh
(2017)
Development and applications of novel approaches for monitoring antibody generation to prostate cancer-related antigens.
PhD thesis, Dublin City University.
Antibody–based approaches have provided vast improvements in the detection and
treatment of many cancers. However, progress is currently limited in prostate cancer
due to a lack of clinically-relevant biomarkers. Novel biomarker discovery may be
achieved with antibody technology, which utilises cancer cells to pan antibody
libraries/sources, to identify antibodies against non-predetermined antigens. The major
limitation currently encountered is the difficulty in establishing the identities of such
antigens, due to the need for extensive post-selection antibody characterisation.
The work embodied in this thesis utilised protein array technology to monitor the
generation of a large recombinant antibody library directed towards nonpredetermined antigens present in a prostate cancer cell line. Protein arrays were used
to profile the immune response, mounted by the avian host following cancer cell
immunisation. Investigations of the antigen profile revealed a trend in the immune
response towards orthologs that were conserved between human and chicken. Similar
protein array-profiling was utilised to characterise the resulting avian scFv-phage library
generated, as well as 384 scFv isolated following enrichment of the library towards
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prostate cancer cells. Evaluation of identified scFv-antigens allowed for the selection of
an antigen subset to be screened against the scFv library. This screening resulted in the
isolation of the first recombinant antibody to Zinc Finger Protein 358, an antigen
currently uncharacterised in prostate cancer. This antibody was isolated and
characterised.
In conclusion, the incorporation of protein array technology into recombinant antibodybased biomarker discovery enabled antigen characterisation at an earlier stage and on
a larger scale than possible with traditional methods, thus circumventing the bottleneck
associated with the current strategies in biomarker discovery using recombinant
antibody technology. The high-throughput approach developed in this study sheds light
on our understanding of immunisation processes and substantially enhances the
potential of recombinant antibody technology for cancer biomarker discovery and
antibody development.
Item Type:
Thesis (PhD)
Date of Award:
November 2017
Refereed:
No
Supervisor(s):
O'Kennedy, Richard and Kijanka, Gregor
Uncontrolled Keywords:
protein array technology; scFv-antigens; avian scFv-phage; 384 scFv