Superparamagnetic nanoparticles containing chelating ligands have gained significant attention in biomedical engineering because of their potential to realize targeted drug delivery, diagnostic imaging, and hyperthermia therapy. In this study, we develop the development of chelate ligand-attached superparamagnetic nanoparticles using nitrilotriacetic acid (NTA) and maghemite (γ-Fe2O3) as key components. To smoothly attach NTA to the surface of γ-Fe2O3 nanoparticles, amino functionalization was performed using polyethyleneimine (PEI) and (3-Aminopropyl) triethoxysilane (APTES), which form easily to silane and polymer structures at the oxide surface and have amine groups at the end, as the polymeric and inorganic precursors, respectively. The surface characteristics of nanoparticles were analyzed in terms of surface charge, while focusing on the effects of amino functionalization and NTA attachment, and the behavior of NTA conjunction through peptide bonds. This study provides an examination of the physicochemical property changes, such as magnetism, surface charge, and morphological changes, that colloidal particles undergo due to property changes caused by the attachment of chelating ligands. Display omitted •γ-Fe2O3-NTA nanoparticles were fabricated through amino functionalization.•(3-Aminopropyl) triethoxysilane (APTES) and polyethyleneimine (PEI) were used as precursors for γ-Fe2O3 surface modification.•NTA was successfully conjugated to γ-Fe2O3 nanoparticles via cross-linking.•The morphology of NTA attachment was depended on the type of amine precursors.