A device for the generation of nanoaerosols from biological substances was designed and the resulting nanoaerosols were characterized. The function of the generator is based on the electrospray-neutralization process in which nanoclusters, obtained by electrohydrodynamic atomization, are neutralized by counter-ions generated by the electrospraying of ethanol at the opposite potential. The device is capable of generating nanoaerosol from suspensions of nanoparticles and solutions of non-volatile organic substances (e.g., antibiotics), proteins, and other biomolecules. This device produces nanoaerosols (10-300 nm in size) with a concentration up to 2 x 10(7) cm(-3). A quartz crystal microbalance based device was designed to monitor the performance of the generator. Using magnetic Fe3O4 nanopartides as reporters, it has been demonstrated that approximately 0.4 mu g of nanoaerosol was deposited into a mouse lung in 1 h in a nose-only exposure system. Thus, we present an approach to both generate nanoaerosol from biological and therapeutic substances and to estimate the doses delivered to mice exposed to these nanoaersolized substances. (C) 2013 Elsevier Ltd. All rights reserved.