Abstract: The aim of this study was to prepare albumin nanoparticles by thermal driven self-assembly, and to investigate the formation mechanism, cellular uptake, the kinetics of cellular uptake and intracellular degradation, etc. By measuring the concentrations of thiol, amino and carboxyl groups, the formation mechanism of albumin nanoparticles was revealed. CCK-8 assay was performed to detect the cytotoxicity; inverted fluorescence microscope was used to observe the cellular uptake of the nanoparticles; while the fluorescence resonance energy transfer(FRET)method was applied to investigate the cellular uptake and intracellular degradation kinetics. The drug-loading capacity was investigated using paclitaxel(PTX)as the model drug. The results showed that the albumin nanoparticles produced by thermal driven self-assembly were safe, nontoxic, biodegradable and stabilized by intermolecular disulfide and amide bonds. The drug-loading study indicates that PTX can be highly encapsulated in the nanoparticles. Hence, thermal driven self-assembly method is green and easy to operate, and the albumin nanoparticles can be applied as a new delivery platform for anticancer drugs.