During the preparation and development of selenium nanoparticles (SeNPs), natural bioactive peptides are added to enhance their physicochemical characteristics and functional properties. Among these properties, immunomodulatory activities, which include activating immune cells to strengthen immunity, constitute the major functions of the immune system. To obtain SeNPs with enhanced immunomodulation, ovalbumin peptide (OP) was used as a stabilizer, yielding OP-SeNPs. The physicochemical properties of OP-SeNPs were characterized. RAW264.7 macrophages were used as a model to investigate the immunomodulatory activity of OP-SeNPs. Results indicate that OP-SeNPs were zero-valent and amorphous, with a particle size of 82.23 ± 1.77 nm. SeNPs demonstrated positive interactions with the -OH, CO, CN, and NH groups of OP. In addition, OP-SeNPs activated RAW264.7 macrophages by increasing NO secretion and enhancing pinocytosis activity, indicating their ability to enhance immunomodulatory effects. Therefore, this study provides a theoretical basis for the construction and characterization of bioactive peptides and SeNP complexes.