Antibacterial polycaprolactone (PCL) electrospun fiber mats were prepared by coelectrospinning PCL with soluble eggshell membrane protein (SEP) in 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP), followed by adsorption of silver nanoparticles (Ag NPs) through hydrogen-bonding interaction between the amide groups of SEP and the carboxylic acid groups capped on the surfaces of Ag NPs. The PCL/SEP fiber mat was characterized by X-ray photoelectron spectroscopy, indicating the presence of some SEP on the fiber surface. The adsorption of Ag NPs was confirmed by transmission electron microscopy and quantitatively characterized by thermogravimetric analysis. The pH value of the silver sol used for adsorption is very important in view of the amount and dispersion state of Ag NPs adsorbed on the fibers. The Ag NP-decorated PCL/SEP fiber mats prepared at pH 3-5 exhibit strong antibacterial activity against both gram-negative Escherichia coli and gram-positive Bacillus subtilis. Antibacterial PCL fiber mats were also obtained similarly with the assistance of collagen (another protein) instead of SEP, showing that protein-assisted adsorption of Ag NPs is a versatile method to prepare antibacterial electrospun fiber mats.