Fluorescent protein nanoparticles: Synthesis and recognition of cellular oxidation damage

Intracellular reactive oxygen species (ROS) generation are associated with many diseases. Lots of studies focus on the detection of intracellular ROS by small fluorescent molecules. However, ROS recognized by biocompatible nanoparticles are relatively less reported. It is widely known that albumin-based nanomaterials possess unique advantages in biomedical applications because they are biodegradable and biocompatible. Herein, fluorescent protein nanoparticles (PNPs) were prepared using BSA as a starting material without introducing extra fluorescent molecules. The blue fluorescent PNPs were well characterized by FL, FTIR, CD, TEM, DLS, etc. It was revealed that the PNPs exhibited two types of emissive centers through FL spectra and the fluorescence lifetimes. Further mechanism study indicated that the fluorescence of the PNPs was mainly derived from three kinds of aromatic amino acids, namely tryptophan, tyrosine and phenylalanine. Moreover, the fluorescence properties of the PNPs were tightly related to pH. The PNPs displayed excellent stabilities under harsh conditions as well as physiological conditions. In addition, the PNPs (200 μg/mL) were nontoxic to HeLa and GES-1 cell lines, showing good biocompatibility. The cellular uptake of PNPs was occurred only when the cells were stressed with glucose oxidase or H₂O₂, thereafter the bright blue fluorescence was observed, indicating that it could be utilized for the recognition of cellular oxidation damage. These findings will offer novel clues for the future synthesis of even brighter protein nanoparticles and their biomedical applications.