Amal Sarhan

I am a PhD researcher in protein and structural biology, specializing in membrane proteins and protein–protein interactions associated with lipid transport in Mycobacterium tuberculosis. Particularly, my research involves rational construct design, genetic engineering, and molecular cloning of membrane protein variants, followed by recombinant expression in E. coli. I performed protein purification using Ni-NTA, Strep-tag, nanoGFP-based affinity, SEC-FPLC, and generated detergent-free and soluble protein preparations. I conducted biophysical characterization using circular dichroism (CD), NanoDSF, and SEC-MALS to assess protein folding, stability, oligomerization state, and sample quality. I investigated protein–protein interactions using the two-hybrid assay in Mycobacterium smegmatis and bimolecular fluorescence in E. coli. I performed structural studies through SAXS data collection and analysis, crystallization screening, and negative stain EM sample preparation.

My master’s research focused on recombinant subunit vaccine development targeting multidrug-resistant bacterial pathogens. The project involved molecular cloning and expression of three novel antigens in E. coli, followed by purification using Ni-NTA affinity chromatography. Purified antigens were applied in murine immunization and challenge models, with vaccine efficacy assessed through bacterial bioburden quantification and histopathological analysis. Antigen-specific immune responses were quantified using indirect and whole-cell ELISA, including IgG subclass profiling to evaluate humoral and cell-mediated immune activation.