Aptamers are small DNA ligands that have been manually selected to strongly and
specifically bind a target of interest. These molecules may prove superior to modern
antibodies in a number of ways including price and reproducibility. One of the major
advantages of using aptamers as opposed to antibodies is the relative speed of
development. This, coupled with the repetitive nature of aptamer selection, means that
the entire process is a possible target for automation. In the following experiments, a
ssDNA aptamer is developed against the human red blood cell protein glycophorin A,
partially through the novel use of a robotized benchtop. The process also utilizes an
adapted protocol for emulsion PCR to further increase the efficiency of the selection
process. After 11 rounds of selection, the DNA pools were sequenced leading to the
generation of 14 potential aptamers. These aptamers were tested with the isolated
protein and with human red blood cells resulting in several of the aptamers being
deemed potential binders. Further work with these identified sequences could result in
aptamers that can be reliably used to tag and delicately separate red blood cells from
other cells of interest within blood, such as stem cells. The novel approaches to
selection used in this work may also lead to quicker and more efficient generation of
future aptamers.
| Identifer | oai:union.ndltd.org:uottawa.ca/oai:ruor.uottawa.ca:10393/38175 |
| Date | 24 September 2018 |
| Creators | Bushnik, Evan |
| Contributors | Berezovski, Maxim |
| Publisher | Université d'Ottawa / University of Ottawa |
| Source Sets | Université d’Ottawa |
| Language | English |
| Detected Language | English |
| Type | Thesis |
| Format | application/pdf |
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