Hemoperfusion is an important strategy for liver disease treatment. Polystyrene-divinylbenzene (PS-DVB) microspheres are widely applied as absorbents in hemoperfusion to efficiently remove the important toxin bilirubin. However, as another common toxin, endotoxin will remain during this process and cause endotoxemia. Therefore, simultaneous removal of both bilirubin and endotoxin is highly desirable. In the present study, we engineered PS-DVB microspheres with polymyxin B sulfate (PMB) to meet this goal. After modification, the novel PMB-engineered (P-PMB) microspheres displayed excellent biocompatibility and hemocompatibility. Notably, compared to PS-DVB microspheres, P-PMB microspheres exhibited markedly stronger detoxification of both bilirubin and endotoxin, increasing by 17.03% and 42.57%, respectively. Overall, we believe that the novel P-PMB microspheres have considerable potential for liver disease treatment in clinical practice.