• protein;
  • self-assembly;
  • cysteine;
  • dithiothreitol;
  • particles


In this article, a non-chemical crosslinking method is used to produce pure protein microparticles with an innovative approach, so-called protein activation spontaneous and self-assembly (PASS). The fabrication of protein microparticles is based on the idea of using the internal disulfide bridges within protein molecules as molecular linkers to assemble protein molecules into a microparticle form. The assembly process is triggered by an activating reagent–dithiothreitol (DTT), which only involved in the intermediate step without being incorporated into the resulting protein microparticles. Conventional protein microparticle fabrication methods usually involve emulsification process and chemical crosslink reactions using amine reactive reagents such as glutaraldehdye or EDC/NHS. The resulting protein microparticles are usually having various size distributions. Most importantly crosslinking reactions using amine reactive reagents will result in producing protein microparticles with undesired properties such as auto-fluorescence and high toxicity. In contrast to the conventional methods, our technology provides a simple and robust method to produce highly homogeneous, stable and non-fluorescence pure protein microparticles under mild conditions at physiological pH and temperature. The protein microparticles are found to be biodegradable, non-toxic to MDCK cells and with preserved biological activities. Results on the cytotoxcity study and enzyme function demonstrate the potential applications of the protein microparticles in the area of pharmaceutics and analytical chemistry.