This paper reports layer-by-layer films fabricated with hemoglobin (Hb) and gold nanoparticles (AuNPs) as well as their applications in carrying oxygen. The characterization of {Hb/AuNPs}(n) multilayer films at different layers revealed that the formation of films was step-by-step. Meanwhile, the study showed that the {Hb/AuNPs}(n) modified glass carbon electrode can maintain the biological activity of Hb. The capacity of carrying oxygen was associated with the number of assembled layers. The {Hb/AuNPs}(3) multilayer films displayed a good storing and releasing ability of oxygen. The study can provide an alternative method for construction of hemoglobin function interface with high-efficiency oxygen carrying capacity. (C) 2015 Elsevier B.V. All rights reserved.