UC Riverside

Engineered nanomaterials (ENMs) have great application potentials in biological systems, while the protein corona formed around ENMs after they encounter any biofluids makes their fate difficult to be predicted. Protein corona provides a brand new “biological identify” for ENMs, which will determine the recognition, targeting, and compatibility of ENMs in vivo. One important property of protein corona is its dynamic exchange of components during incubation time, on which lots of endeavors have been made, but there is still knowledge gap between the intrinsic properties of ENMs and what kinds of protein will form the corona. Moreover, the molecular details of protein in corona, including the orientation, conformational change and aggregation, are also important for the function of protein corona. Due to the non-specific forces behind protein-ENMs interactions, desired and controlled arrangement of protein on ENMs surface has always been a significant but difficult task. Despite of various success made on those two topics, rapid and routine methods for composition analysis and molecular details exploration of protein corona are still in stark deficient. This research will focus on new methods development for those two problems. Firstly, a fluorescamine labeling based high throughput screening method is applied to screen and discriminate interactions between single protein and ENMs, which could indicate either protein binding or unfolding induced by ENMs. With those results as descriptors, correlations between them and protein corona composition have been found, which suggests that a structure activity quantification model using those descriptors could be built for rapid corona prediction. Secondly, limited proteolysis coupled with LC-MS/MS capable to identify binding sites of protein on another molecule has been developed. By applying it to those positive protein-ENMs pairs identified in previous screening, the molecular details including orientation and unfolding of proteins in corona could be unveiled. Despite of the limitation on precision, information obtained from this method could be helpful for further rational design of ENMs on biological application.