Electrospun nanofiber surfaces are modified with proteins to control protein release. A mixture of poly(E-caprolactone) (PCL) and PCL-PEG block copolymers is electrospun to prepare amine-terminated block copolymers. The amount of surface exposed amine groups increases as the blend ratio of block copolymer increases. Cell attachments on the nanofibers change according to the ratio of the block copolymer in the blend; this indicates that the PEG moiety plays a significant role in enhancing and decreasing biocompatibility of nanofibers. Fluorescent proteins are immobilized on the surface of nanofibers by conjugating activated carboxylic groups of the protein to the surface exposed amine groups of nanofibers in an aqueous environment. The number of amine groups increases as the ratio of block copolymers increases. Proteins that are chemically associated with the nanofibers show an attenuated release profile while most of the proteins physically associated with the nanofibers are released in 1 day. These results show that the protein-immobilized nanofibers can be potentially applied to tissue engineering scaffolds and wound healing materials with bioactive protein being slowly released over a long period of time.