Two synthetic human antibody fragments against the human pentaspan membrane protein CD133 were isolated using a novel selection method involving direct selections on cells coupled with Illumina deep sequencing. The antibody fragments were isolated through a PCR-based recovery strategy developed in the lab and subsequently converted to full length IgGs. Termed RW01 and RW03, the antibodies bind separate epitopes on CD133 and are able to detect the protein using various molecular techniques. Finally, experiments have shown that RW01 and RW03 treatment affect the stability of CD133 on live cells. Additional experiments are required to reveal the specific epitope recognized by each antibody, which organelles they are targeted to when internalized and whether they have an effect on cellular differentiation or cellular viability. In addition to the therapeutic potential of these antibodies, they will have many applications towards expanding our knowledge concerning the CD133 protein and its role in cancer.
| Identifer | oai:union.ndltd.org:TORONTO/oai:tspace.library.utoronto.ca:1807/43333 |
| Date | 11 December 2013 |
| Creators | Williams, Rashida |
| Contributors | Moffat, Jason, Sidhu, Sachdev |
| Source Sets | University of Toronto |
| Language | en_ca |
| Detected Language | English |
| Type | Thesis |
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