Construction of nanoscale protein particle using temperature-sensitive elastin-like peptide and polyaspartic acid...

Temperature-responsive monodisperse spheres are useful for various in vivo and in vitro applications. Size, response temperature and biocompatibility are particularly important consideration with in vivo applications. In this work, we constructed fusion proteins of low antigenic elastin-like peptide (ELP) and a polyaspartic acid chain, and studied the particles that had a favorable size and temperature of formation of particle. From DLS analysis, we confirmed that some of them formed particles with less than 100nm in diameter around 37 degrees C, while the diameter of ELPs alone is larger than 1microm in diameter. The (PGVGV)(160)D(22), which is composed of a short aspartic acid chain and a long ELP region, had a tendency to form large particles. The temperature of formation and collapse of the protein particle were dependent on the length of the ELP and the polyaspartic acid chain, and the concentration of proteins. The direct observation with TEM indicated that the morphologies of the particles were spherical except when (PGVGV)(160)D(22) was used. The intensities of the environment-sensitive hydrophobic fluorescence increased at 37 degrees C more than 1.5 times as much as at 25 degrees C both in free form and modified at the ELP region. These results indicated that the polarity of the environment surround the fluorescence decreased or the movement of fluorescence was limited, and thus, implied that the ELP formed a more hydrophobic or rigid region and could hold hydrophobic drugs. These results suggest that a temperature-responsive protein particle with favorable size and temperature of formation can be constructed that is suitable for any in vitro or in vivo application.