Objective To establish an enzyme linked immunoassay (ELISA) for human bone alkaline phosphatase (BAP). Methods A double antibody sandwich method was used to develop the ELISA method, in which one strain was coated on the microplate and another strain was labeled with horseradish peroxidase (HPR). TMB color was used for chromogenic system. Results The BAP detection range was from 5 to 120ng/ml, the correlation coefficient of the standard curve was more than 0.995, and the lowest detection level was 1.5ng/ml. The average recovery rate was between 96.4% and 102%, the intraassay CV was between 5.6% and 7.9%, and the BAP normal concentration in serum of healthy men was less than 10ng/ml. 502 serum samples were tested by this method, and the results of the ELISA corresponded to those of the control kits. Conclusion the ELISA developed in this study might be suitable for bone alkaline phosphatase analysis.