Hyperbilirubinemia is one of most severe clinical diseases, which is caused by the accumulation of unconjugated bilirubin. Electrospun nanofiber membranes used as highly efficient bilirubin adsorbents have been applied to remove the extra bilirubin in hemoperfusion for their high surface area and easy functionalized properties. In this work, Lysine (Lys) grafted polyacrylonitrile (PAN) electrospun nanofiber membranes doped with organic Hectorite (OHec) (Lys-HPAN@OHec) have been fabricated via a series of modified process, including pore forming, alkaline hydrolysis and grafting reaction. The obtained Lys-HPAN@OHec nanofiber membranes have been analyzed in detail and investigated the adsorption capacity for bilirubin. Compared with original PNA membranes, Lys-HPAN@OHec nanofiber membranes show an excellent bilirubin adsorption capacity and more stable rejection rate of bovine serum albumin (BSA). The maximum adsorption capacity of Lys-HPAN@OHec membranes for bilirubin is 64 mg/g, the adsorption process of Lys-HPAN@OHec membranes matched the Langmuir model well. In addition, dynamic adsorption reveals that the adsorption equilibrium time of Lys-HPAN@OHec membranes is about 2 h. Significantly, Lys-HPAN@OHec membranes have excellent biocompatibility and hemocompatibility. This study demonstrates that the novel Lys-HPAN@OHec membranes may provide a new way to treat hyperbilirubinemia.