The present study mainly compared the effect of docetaxel-albumin nanoparticles (DANPs) and docetaxel-loaded PEG-albumin nanoparticles (PEG-DANPs) against non-small cell lung cancer (NSCLC). We made systematic assessments on these three drugs against NSCLC both in vitro and in vivo. With the purpose of eliminating side-effects of the commercial formulation (Tween-80) and prolonging the blood circulation time, we used emulsion-evaporation cross-link method to prepare DANPs and PEG-DANPs. The DANPs had an average particle size of 163.4 ± 3.76 nm, a zeta potential of -19.4 ± 0.18 mV, a polydispersity index of 0.143 ± 0.03, a drug loading of 8.71 ± 0.98%, and an encapsulation efficiency of 93.58 ± 0.86%; the average particle size of PEG-DANPs is 169.19 ± 2.36 nm, zeta potential is -18.2 ± 0.21 mV, with a polydispersity index of 1.56 ± 0.05, a drug loading of 8.72 ± 1.05% and an encapsulation efficiency of 95.4 ± 5.5%. PEG-DANPs showed a dose- and time-dependent efficacy in cytotoxicity studies in vitro; the hemolysis test indicated that PEG-DANPs had less hemocytolysis than Aisu® and DANPs; in addition, a more prolonged circulation time and sustained in vitro release behavior were observed in the PEG-DANPs compared with Aisu® and DANPs; the cellular uptake test in vitro demonstrated that PEG-DANPs could be absorbed easier into the nucleus; furthermore, the tumor growth of NSCLC-bearing nude mice in vivo was reduced the most by PEG-DANPs. In conclusion, the PEG-DANPs have the lowest side-effects, the highest antitumor activity with the longest blood circulation time of the three drugs, and it will provide an alternative to patients with NSCLC.