Department of Biochemistry, Faculty of Science, King Abdulaziz University (KAU), Jeddah, Saudi Arabia; Institute for Synthetic Bioarchitectures, Department of Nanobiotechnology, University of Natural Resources and Life Sciences (BOKU), Vienna, Austria. Electronic address: [Email]
Development of synthetic bioarchitectures to improve our understanding of biological systems and produce biosynthetic models with new functions has attracted substantial interest. Synthetic HDL-like phospholipid nanodiscs are a relatively new model of nanoparticles that present a promising carrier for drug delivery and membrane protein investigations. Nanodiscs are soluble nanoscale phospholipid bilayers that are produced based on the self-assembly of phospholipids, membrane scaffold proteins (MSP) and an embedded peptide/protein of interest. To determine the effect of conjugating a protein with a probe, the model protein bovine serum albumin (BSA) with or without FITC conjugation was attached onto 100% 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-choline (POPC) nanodiscs. The generated discs were analyzed by Fast Protein Liquid Chromatography (FPLC), dynamic light scattering (DLS), and UV-VIS spectroscopy. Empty, BSA- and FITC-BSA-Nanodiscs exhibited different size, charge and elution characteristics as well as different release profiles. Thus, conjugation of proteins to be adsorbed onto nanodiscs surfaces with fluorophores can affect the physical and release properties of nanodiscs, thereby potentially impacting their biophysical, delivery and imaging applications.