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Since chelating chemical had focused for enhancing bio-susceptibility and ability for the binding with biomaterials, in thisstudy, Nitrilotiriacetic acid (NTA) was utilized for the developing chelate ligand attached superparamagnetic nanoparticle. Inorder to conjugate NTA, carboxylic group was needed on the surface of substrate particle for building peptide bond withamine group which was branched from the NTA structure. Therefore Fe3O4 nanoparticle was functionalized with various ofamine precursor (Polyethyleneimine (PEI), (3-Aminopropyl)triethoxysilane (APTES)), and carboxyl precursor (Polyacrylicacid (PAA)) before the conjugation of NTA. Then NTA conjugation behavior was estimated which is varied with the type ofamine precursor which affects to the surface properties.