In head and neck oncology, the oncogenic viral role is currently debated. Particularly, the infection by Human Papillomavirus (HPV) demonstrated oncogenic properties in the development of oropharyngeal cancers. Nowadays there are no recommended or validated methods to screen for early cases of HPV-driven oropharyngeal cancer. The present work developed and tested in vitro and in vivo performance of an innovative methods to early detect HPV-positive (HPV+) oropharyngeal squamous cell carcinoma (OPSCC). This system is represented by a biosensor for the detection of HPV antibodies against HPV E6 and E7 oncoproteins using the Electrochemical Impedance Spectroscopy (EIS) technique. Assays to quantify HPV-specific antibodies were based on Enzyme Linked Immuno-Sorbent Assays (ELISA); ELISA requires specialized laboratory and personnel, it is quite expensive, complex and time-consuming. Moreover, it requires pre-steps of sample processing, that can cause proteins denaturation and bioactivity modification. An instantaneous self-test would be particularly useful to detect the presence of HPV antibodies in blood, both for diagnostic and prognostic purposes. A cheap test could really impact on developing countries in the context of cancer prevention. The work is composed of three parts: 1. development of a biosensor based on gold screen-printed electrode (Au-SPE) functionalized with self-assembled monolayer (SAM) s and a commercial mixture of E7 peptides (PepMix) acting as the antigen to capture serum antibodies; 2. the second part of the work was focused on the recombinant production of E6 oncoproteins and a different electrode functionalization using hybrid SAMs. 3. the last part reported the analysis of biological samples testing the device in different conditions.
APPLICATION OF NANOTECHNOLOGY TO THE DIAGNOSIS AND TREATMENT OF ORAL AND OROPHARYNGEAL CANCER
TOFANELLI, MARGHERITA
2020
Abstract
In head and neck oncology, the oncogenic viral role is currently debated. Particularly, the infection by Human Papillomavirus (HPV) demonstrated oncogenic properties in the development of oropharyngeal cancers. Nowadays there are no recommended or validated methods to screen for early cases of HPV-driven oropharyngeal cancer. The present work developed and tested in vitro and in vivo performance of an innovative methods to early detect HPV-positive (HPV+) oropharyngeal squamous cell carcinoma (OPSCC). This system is represented by a biosensor for the detection of HPV antibodies against HPV E6 and E7 oncoproteins using the Electrochemical Impedance Spectroscopy (EIS) technique. Assays to quantify HPV-specific antibodies were based on Enzyme Linked Immuno-Sorbent Assays (ELISA); ELISA requires specialized laboratory and personnel, it is quite expensive, complex and time-consuming. Moreover, it requires pre-steps of sample processing, that can cause proteins denaturation and bioactivity modification. An instantaneous self-test would be particularly useful to detect the presence of HPV antibodies in blood, both for diagnostic and prognostic purposes. A cheap test could really impact on developing countries in the context of cancer prevention. The work is composed of three parts: 1. development of a biosensor based on gold screen-printed electrode (Au-SPE) functionalized with self-assembled monolayer (SAM) s and a commercial mixture of E7 peptides (PepMix) acting as the antigen to capture serum antibodies; 2. the second part of the work was focused on the recombinant production of E6 oncoproteins and a different electrode functionalization using hybrid SAMs. 3. the last part reported the analysis of biological samples testing the device in different conditions.File | Dimensione | Formato | |
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https://hdl.handle.net/20.500.14242/62611
URN:NBN:IT:UNITS-62611