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The development of an electrochemical biosensor to detect and measure autophagy flux
Thesis (MEng) -- Stellenbosch University, 2022.
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| Format: | Thesis |
| Language: | en_ZA |
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Stellenbosch : Stellenbosch University
2022
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| _version_ | 1867614037977595904 |
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| access_status_str | Open Access |
| author | Jacobs, Kenneth |
| author2 | Perold, Willem |
| author_browse | Jacobs, Kenneth Perold, Willem |
| author_facet | Perold, Willem Jacobs, Kenneth |
| author_sort | Jacobs, Kenneth |
| collection | Thesis |
| dc_rights_str_mv | Stellenbosch University |
| description | Thesis (MEng) -- Stellenbosch University, 2022. |
| format | Thesis |
| id | oai:scholar.sun.ac.za:10019.1/125972 |
| institution | Stellenbosch University (South Africa) |
| language | en_ZA |
| last_indexed | 2026-06-10T12:45:40.774Z |
| license_str | Other — see source repository |
| provenance_str_mv | Harvested via OAI-PMH from SUNScholar — Stellenbosch University Repository |
| publishDate | 2022 |
| publishDateRange | 2022 |
| publishDateSort | 2022 |
| publisher | Stellenbosch : Stellenbosch University |
| publisherStr | Stellenbosch : Stellenbosch University |
| record_format | dspace |
| source_str | SUNScholar — Stellenbosch University Repository |
| spelling | oai:scholar.sun.ac.za:10019.1/125972 The development of an electrochemical biosensor to detect and measure autophagy flux Jacobs, Kenneth Perold, Willem Stellenbosch University. Faculty of Engineering. Dept. of Electrical and Electronic Engineering. Autophagic vacuoles Biosensors Electrochemical analysis Biochemical markers UCTD Thesis (MEng) -- Stellenbosch University, 2022. ENGLISH ABSTRACT: Autophagy is a cellular process, which a cell undergoes self-degradation to remove unwanted and dysfunctional cellular material. The autophagic process is associated with various diseases such as cancers, neurodegenerative, and infectious diseases, as well as longevity and health. The protein LC3-II is the most common biomarker used to detect the autophagic process. LC3-II is directly related to the formation of autophagosomes, a double-membraned vesicle that contains cellular material for degradation, therefore the change in the concentration of LC3-II is a good estimate of autophagy flux. A label-free electrochemical DNA biosensor is developed to measure the concentration of the DNA that encodes the protein LC3-II. Through covalent immobilization of thiol modified ssDNA probes onto a planar gold electrode, a self-assembled monolayer of ssDNA probes is formed, which acts as the integrated transducer and recognition layer. The ssDNA targets are detected through the hybridization reaction of complementary ssDNA targets and ssDNA probes. A low cost potentiostat is developed to apply square-wave voltammetry to the sensor, which is submerged in an electrolyte solution of potassium ferrocyanide in potassium chloride. The hybridization reaction results in a change in the electrical properties of the sensor which is exploited to measure the concentration of the ssDNA targets. The results indicated that the decreasing peak current measured by the biosensor is directly related to the increasing concentration of the ssDNA targets. AFRIKAANS OPSOMMING: Outofagie is ’n sellulêre proses waar selle selfdegradasie ondergaan om ongewenste en disfunksionele sellulêre materiaal te verwyder. Die outofagiese proses word geassosieer met kanker, neurodegeneratiewe, aansteeklike siektes, sowel as langlewendheid en gesondheid. Die proteïen LC3-II is die mees algemene biomerker wat gebruik word om die outofagiese proses op te spoor. LC3-II is direk verwant aan die vorming van outofagosome, ’n dubbelmembraan vesikel wat sellulêre materiaal vir degradasie bevat en daarom is die verandering in die konsentrasie van LC3-II ’n goeie skatting van outofagie vloed. ’n Etiketvrye elektrochemiese DNS biosensor word ontwikkel wat die konsentrasie van die DNS wat die proteïen LC3-II kodeer, meet. Deur kovalente immobilisasie van thiolgemodifiseerde ssDNS-probes op ’n plat goue elektrode, is ’n self-saamgestelde monolaag van ssDNS-probes gevorm, wat dien as die geïntegreerde omsetter en herkenningslaag. Die ssDNS-teikens word opgespoor deur die hibridisasiereaksie van komplementêre ssDNS-teikens en ssDNS-probes. ’n Laekoste potensiostaat word ontwikkel om vierkantgolfvoltammetrie op die sensor toe te pas, wat in ’n elektrolietoplossing van kaliumferrosianied in kaliumchloried lê. Die hibridisasiereaksie verander die elektriese eienskappe van die sensor. Die dalende piekstroom gemeet deur die biosensor hou direk verband met die toenemende konsentrasie van die ssDNS-teikens. Masters 2022-11-16T06:52:35Z 2023-01-16T12:43:44Z 2022-11-16T06:52:35Z 2023-01-16T12:43:44Z 2022-12 Thesis http://hdl.handle.net/10019.1/125972 en_ZA Stellenbosch University xvi, 118 pages : illustrations application/pdf Stellenbosch : Stellenbosch University |
| spellingShingle | Autophagic vacuoles Biosensors Electrochemical analysis Biochemical markers UCTD Jacobs, Kenneth The development of an electrochemical biosensor to detect and measure autophagy flux |
| title | The development of an electrochemical biosensor to detect and measure autophagy flux |
| title_full | The development of an electrochemical biosensor to detect and measure autophagy flux |
| title_fullStr | The development of an electrochemical biosensor to detect and measure autophagy flux |
| title_full_unstemmed | The development of an electrochemical biosensor to detect and measure autophagy flux |
| title_short | The development of an electrochemical biosensor to detect and measure autophagy flux |
| title_sort | development of an electrochemical biosensor to detect and measure autophagy flux |
| topic | Autophagic vacuoles Biosensors Electrochemical analysis Biochemical markers UCTD |
| url | http://hdl.handle.net/10019.1/125972 |
| work_keys_str_mv | AT jacobskenneth thedevelopmentofanelectrochemicalbiosensortodetectandmeasureautophagyflux AT jacobskenneth developmentofanelectrochemicalbiosensortodetectandmeasureautophagyflux |