Moran, Kara (2018) Development of recombinant antibody fragments for mycotoxin detection and the investigation of AFB1 and anti-AFB1 Fab antibody fragment effects in hepatocellular carcinoma. PhD thesis, Dublin City University.
Abstract
Mycotoxins are toxic secondary metabolites produced by fungal species that grow on a vast array of food commodities globally. These contaminants pose a massive concern for human and animal health due to their prevalence in commonly consumed food commodities. Studies have shown that mycotoxin exposure, in particular aflatoxin B1 (AFB1) exposure, imparts mutagenic and carcinogenic effects in humans and animals. Accordingly, research is now focusing on methods to allow the rapid, cost-effective and simple detection of mycotoxins in food samples directly at the site of contamination. Additionally, investigations are centred on an improved understanding of the mechanisms that underpin mycotoxic carcinogenic effects.
This thesis describes the development and optimization of recombinant antibody fragments for inclusion in a ‘point-of-site’ device to allow the sensitive and specific detection of AFB1 and T-2 toxin in the animal feed component, Dried Distillers Grain with Solubles (DDGS). A simple optical-planar waveguide cartridge and detection system that incorporated a recombinant anti-AFB1 Fab antibody fragment and an anti-T-2 toxin polyclonal antibody, in a competitive inhibition format, was used for the successful detection of AFB1 and T-2 toxin, to legislative limits outlined by the European Union. The system also allowed the simultaneous detection of both mycotoxins in a ‘proof-of-concept’ study.
This research also focused on investigating the effects of AFB1 on HepG2 human hepatoma cells and hypothesised that the anti-AFB1 Fab antibody fragment may have a cognate ligand on HepG2 cells. The findings outlined herein, demonstrate that low doses of AFB1 have an inhibitory effect on HepG2 cell migration and proliferation after an initial 24 hours of exposure followed by increased section of IL-8 and recovery of the cells to normal levels of migration and proliferation. In addition, these results established that the anti-AFB1 Fab antibody fragment interacted with, and, exerted an inhibitory effect on hepatocellular carcinoma cell migration and proliferation.
Metadata
Item Type: | Thesis (PhD) |
---|---|
Date of Award: | November 2018 |
Refereed: | No |
Supervisor(s): | O'Kennedy, Richard, O'Neill, Sandra and O'Reilly, Julie-Ann |
Subjects: | Biological Sciences > Biotechnology Biological Sciences > Food technology Biological Sciences > Biosensors |
DCU Faculties and Centres: | DCU Faculties and Schools > Faculty of Science and Health > School of Biotechnology |
Use License: | This item is licensed under a Creative Commons Attribution-NonCommercial-No Derivative Works 3.0 License. View License |
Funders: | Daniel O'Hare Scholarship |
ID Code: | 22361 |
Deposited On: | 22 Nov 2018 16:23 by Julie Ann O'Reilly . Last Modified 17 May 2020 03:30 |
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