Here, the bio-nanocomposite (n-HFP + n-HFS)@An was developed to simultaneously immobilize Pb, Cd and As in the severely contaminated soil. The immobilization rates of diethylenetriaminepentaacetic acid (DTPA)/decarbonate-extracted bioavailable Pb, Cd and As were 59.87%, 31.28% and 62.30%, and the immobilization rates of their water-soluble forms were 63.12%, 60.02% and 89.39%, respectively. Moreover, the ten-year acid rain simulated leaching assay showed that the maximum cumulative release contents of Pb, Cd and As in the treated soil samples were decreased by 2.94, 2.46 and 40.60 times, comparing to the un-treated ones. Additionally, the results of SBRC (Solubility Bioaccessibility Research Consortium) revealed that the bioaccessible rates of the three metals in intestinal phase were lower than in gastric phase, and both of them decreased with increasing the immobilization time. The gastric bioaccessibility of Pb, Cd and As had a higher correlation with the contents of water-soluble forms, while the intestinal bioaccessibility was more strongly positively associated with the bioavailable forms. (C) 2020 Elsevier Ltd. All rights reserved.