Unravelling the molecular armor, cellular dynamics and nuclear trafficking of ultra-stable oligopeptylated-AuNPs: PEG-rivalling nanocargos

Ultra-stable and biocompatible gold nanoparticles (AuNPs) are essential for targeted nanomedicine, facilitating extended circulation, minimal immunogenicity, and efficient cellular uptake. Despite its status as a gold standard for attaining biocompatibility and stability, polyethylene glycol (PEG) faces increasing scrutiny due to its physiology-associated accelerated blood clearance, immunogenicity, and restricted nuclear access, prompting an urgent shift toward alternative surface engineering strategies. This is an extension of our previous study where we fabricated ultra-stable AuNPs using base-etched fish scales, exhibiting PEG-Au comparable physicochemical, mechanical and biofluidic stability. Herein, through integrated surface characterization studies, MALDI-TOF, LC-MS/MS and bioinformatics profiling, we elucidate the shielding oligopeptide consortium that modulates the particles' biomolecular interactions while preserving biofluidic integrity and colloidal stability. Cytotoxicity assays and mechanistic studies of cellular uptake confirmed that the oligopeptylated-AuNPs are non-toxic and are endocytosed <i>via</i> clathrin- and scavenger-mediated receptors. Remarkably, ultra-microtome-assisted HR-TEM revealed that our nanocargos could successfully get imported into the nucleus, a rare and highly significant phenomenon, for such non-viral delivery systems. Collectively, our findings position our sustainably bioengineered oligopeptylated-AuNPs as next-generation nanocargos that uniquely integrate biocompatibility, stealth properties and nuclear-targeting capability, offering a versatile and promising platform to enable precision delivery of therapeutic payloads at subcellular resolutions.