Top-down Nanoparticle-corona by cIEF-MS/MS

In the rapidly evolving field of nanomedicine, understanding the interactions between nanoparticles and biological systems is crucial. A pivotal aspect of these interactions is the formation of a protein corona when nanoparticles are exposed to biological fluids, which significantly influences their behavior and functionality. This study introduces an advanced capillary isoelectric focusing-tandem mass spectrometry (cIEF-MS/MS) platform designed to enhance top-down proteomic analysis of nanoparticle protein coronas. By employing a novel 'sandwich' injection method, our cIEF-MS/MS system significantly reduces analysis time to 30 minutes per run, compared to traditional methods, taking over an hour. This method allows for the rapid, reproducible, and automated profiling of complex protein coronas, identifying over 70 distinct proteoforms per analysis from human plasma-incubated polystyrene nanoparticles. The system’s impressive high-throughput capability and precision in detecting post-translational modifications provide deep insights into the proteomic landscape, unveiling potential biomarkers for various diseases. Our findings augment the understanding of nanoparticle-protein interactions and open new avenues for developing targeted nanomedicine drug delivery systems and diagnostic tools. This study sets a new standard in nanoparticle corona research, promising significant advancements in the field.