Master Degree / Yüksek Lisans Tezleri

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Author: search.filters.author.Taşkent Sezgin, HümeyraPublisher: search.filters.publisher.Izmir Institute of Technology

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  • Master Thesis
    Developing a Lamp Pcr Based Diagnostic Test for Crimean Congo Hemorrhagic Fever Virus
    (Izmir Institute of Technology, 2022) Üstün, Selcen; Taşkent Sezgin, Hümeyra; 01. Izmir Institute of Technology
    Crimean-Congo hemorrhagic fever infection is one of the most common tickborne viral infections, non-infectious in animals and fatal to humans with a mortality rate around 40%. The high mortality rate and lack of vaccines or drug treatment point to the potential danger of infection. The aim of this thesis was to detect 21 complete S-segment Crimean-Congo hemorrhagic fever virus strains originating in Turkey using Fluorimetric Loop Mediated Isothermal Amplification (LAMP) method, which is one of the PCRbased isothermal amplification methods for detecting viral infections. Conserved regions of the 21 strains of the CCHFV observed in Turkey were determined by sequence alignment and LAMP primers for these conserved regions were designed. DNA of CCHFV Ank-2 (GeneBank accession number: MK309333) strain was used and optimum LAMP reaction conditions were determined by changing the temperature, primer amount and MgSO4 concentration. LAMP results were compared with qPCR, which is considered the gold standard. The detection limit for LAMP and qPCR was 2x106 copies/µl. In the study, the CCHFV LAMP primer set-1 gave similar results to the qPCR primer set. The CCHFV LAMP primer set-4 performed better by turning positive 9 minutes earlier than the qPCR primer set. This results indicate LAMP is an alternative method for detecting CCHFV but reveals the necessity of improving the sensitivity of the test.
  • Master Thesis
    Investigation of the Interaction and Olgiomerization of Hiv Capsid and Single Domain Antibody as a Biotechnological Drug Against Hiv
    (Izmir Institute of Technology, 2022) Güney, Seniha; Taşkent Sezgin, Hümeyra; 01. Izmir Institute of Technology
    Human immunodeficiency virus (HIV) causes AIDS which is still a global public health threat. Current drugs against HIV infection cannot eradicate the virus therefore, research on new drug targets continues. HIV capsid protein, which has a highly conserved sequence and is sensitive to mutations, has critical roles in the virus lifecycle, making it a high-potential drug target. A nanobody is the antigen-binding domain of heavy-chain only antibodies of camelids. Small size, thermal stability and ease of production makes nanobodies ideal antibody fragments for therapeutic and diagnostic purposes. In the literature, a nanobody binding to the HIV-1 capsid-N terminal domain (NTD) has been reported. The aim of this thesis is to examine the potential of this nanobody as a biotechnological drug candidate against the HIV-1 and HIV-2 capsid proteins. In the study, HIV-1 capsid was expressed, purified and biophysically characterized. Thermal and chemical denaturation of the protein were done, the melting temperature and unfolding free-energy values of the protein were determined. In-vitro oligomerization of the HIV-1 capsid was performed and observed that the protein self-oligomerized over time. Pure HIV-2 capsid protein could not be produced recombinantly. Thereupon, HIV1 capsid-NTD and HIV-2 capsid-NTD proteins were expressed and purified. Secondary structure of HIV-1 capsid, HIV-1 capsid-NTD and nanobody were analyzed with circular dichroism (CD) spectroscopy and the results matched with the literature. Isothermal titration calorimetry (ITC) experiments were done to examine the HIV-1 capsidnanobody interaction, but good binding was not observed between the two proteins. Future work requires repeating ITC experiments.