I am a PhD holder from the School of Pharmacy, University of Nottingham, where my research was focused on developing novel cellular reporter systems for diagnostic purposes. My research project was, also, emphasis on biochemical engineering, molecular cell biology, gene manipulation and editing strategies, protein biosynthesis and cell culture using a wide range of analytical techniques. The aim of my PhD research project was to study the function and allergenicity of some recombinant parasitic proteins for diagnostic purposes and vaccine development using generated humanised rat basophilic leukaemia (RBL) cell lines, RS-ATL8 NFAT and NPY-mRFP-RBL using two different reporter systems, luciferase and fluorescence, respectively.
I have obtained extensive laboratory experience during my undergraduate, Master and PhD studies. During my PhD, I gained considerable experience in molecular biology approaches, such as PCR techniques, primer designing, mutations, DNA editing and manipulating, amplification, restriction digestion and ligation, cloning genes into different cloning and expression vectors, DNA verification, sequencing, analysing and gene purification. In addition, I have extensive experience in protein synthesis using different expression systems, such as those using cell-free wheat germ lysate, bacterial and mammalian expression systems using human embryonic kidney (HEK293) cell line, and protein purification using different methods of IMAC. This mainly involved expressing and purifying few recombinant Echinococcus and Schistosoma antigens and allergens for screening in the RS-ATL8 NFAT-Luciferase reporter cell line, which utilises IgE cross-linking using plate-reader based assays (immunoluminescence assay). My PhD research project, also, included gene manipulation and editing strategies to overcome the issue of rare codon usage in the expression hosts.
In addition, my work involved gene delivery strategies for developing a novel humanised rat basophilic leukaemia (NPY-mRFP-RBL) reporter system to allow human IgE binding to its own receptor (HsFceRI) on the surface of the rat cells. Fluorescence microscopy, confocal microscopy, fluorescence assay and flow cytometry analysis were utilised to verify the successful transfection.
Moreover, I have a great experience in nucleotides and peptides sequences analysis using different software such as NCBI and SnapGene, which will give a prediction of what is likely to happen. Furthermore, I have great experience in DNA and protein detection and verification using many different detection strategies such as Gel Electrophoresis, SDS-PAGE, Coomassie gel staining, Silver Staining, Western Blot and FUJI-LAS 4000 imaging system. I have also an excellent experience in using optical and fluorescence microscopies, and plate reader for detecting and measuring the luminescence and fluorescence and analysing data.
In my MSc thesis entitled "Pathological Effect of Irradiated Toxoplasma gondii and its Immune Response in Albino Mice", I studied the development of the immune response against toxoplasmosis in Swiss albino mice of the species Mus musculus, BALB/c strain, which were infected, experimentally, with Toxoplasma gondii in order to render the host to resist an infection with this disease in the future. The histopathological changes, humoral immune responses, represented by IgM and IgG; and an innate (non-specific) immune response, represented by phagocytosis were studied in the experimentally infected mice with the parasite Toxoplasma gondii. During my study, animals colonies were grown and maintained. Mice were, also, dissected in the laboratory to harvest tissues and isolate the parasitic cysts from different body organs.
I find that it is valuable to build strong relationships and collaborate with other professional academics at other universities, along with receiving mutual benefits for the team.
