▎ 摘　　要

Hollow TiO2 modified reduced graphene oxide microspheres (hollow TiO2-rGO microspheres or H-TiO2-rGO MS) have been synthesized and then be used to immobilize hemoglobin (Hb) to fabricate a mediator-free biosensor. The morphology and structure of hollow TiO2-rGO microspheres were characterized by scanning, electron microscopy, transmission electronic microscopy and X-ray diffraction. Results of spectroscopy and electrochemistry tests revealed that hollow TiO2-rGO microsphere is an excellent immobilization matrix with biocompatibility for redox protein, affording good protein bioactivity and stability. The hollow TiO2-rGO microspheres with special structure and component enhance the immobilization efficiency of proteins and facilitate the direct electron transfer, which result in the better H2O2 detection performance-the wide linear range of 0.1-360 mu M for H2O2 (sensitivity of 417.6 mu A mM(-1) cm(-2)) and the extremely low detection limit of 10 nM for H2O2. Moreover, the hollow microsphere can provide a protective microenvironment for Hb to make the as-prepared biosensor improve long-term stability. The as-prepared biosensor retains 95.4% of the initial response to H2O2 after 60-d storage. Hence, this work suggests that if can be fabricated a mediator-free biosensor, hollow TiO2-rGO micro spheres will find wide potential applications in environmental analysis and biomedical detection. (C) 2016 Elsevier B.V. All rights reserved.