In this study, a porous silk fibroin (SF) scaffold was modified with soluble eggshell membrane protein (SEP) with the aim of improving the cell affinity properties of the scaffold for tissue regeneration. The pore size and porosity of the prepared scaffold were in the ranges 200–300 μm and 85–90%, respectively. The existence of SEP on the scaffold surface and the structural and thermal stability were confirmed by energy-dispersive X-ray spectroscopy, X-ray diffraction, Fourier transform infrared spectroscopy, differential scanning calorimetry, and thermogravimetric analysis. The cell culture study indicated a significant improvement in the cell adhesion and proliferation of mesenchymal stem cells (MSCs) on the SF scaffold modified with SEP. The cytocompatibility of the SEP-conjugated SF scaffold was confirmed by a 3-(4,5-dimethyltriazol-2-y1)-2,5-diphenyl tetrazolium assay. Thus, this study demonstrated that the biomimic properties of the scaffold could be enhanced by surface modification with SEP. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014, 131, 40138.