Protein affinity membranes based on polyethersulfone (PES) and bovine serum albumin (BSA) were prepared by using an electrospinning method. The changes in the size of nanofiber according to the various conditions of electrospinning (concentration of spinning solution, voltage, flow rate, tip to collector distance) were investigated, and the preparation conditions of the affinity membrane were optimized. XPS and FT-IR results showed the firm incorporation of BSA with PES. It was found PES-BSA nanofibers could be prepared at the temp. of $20{sim}22^{circ}C$ and humidity of $45{sim}55%$. The size of nanofibers increased with increase of the content of PES in the spinning solution while BSA did not affect much to the size of nanofiber. It was also revealed that the narrow size distribution of nanofiber could be obtained with PES 7 wt%, BSA 0.7 wt%, HFP 92.3 wt% at the electrospinning conditions of 1 mL/hr, 10 cm TCD, and 10 kV.