Novel copper metal organic framework nanoparticles Cu-MOF-NPs (C1) were prepared via two simple alternative methods and confirmed by analytical characterization using mass, IR, Raman, XRD spectrum, HR-TEM and TGA-DSC. Mass spectroscopy revealed the molecular ion peak at 647 m/z for the monomeric unit structure n[Cu(AIP)2(PIY)(H2O)2]·4H2O, the presence of which was further supported by mass fragmentation. The Raman spectrum revealed two separate peaks corresponding to D and G bands of carbon in the structure of C1. Moreover, TGA-DSC showed the presence of CuO. XRD data were typically consistent with Raman and TGA-DSC data. In addition, HR-TEM revealed that the morphology of the C1 nanoparticles is uniform with well-distributed elliptical/spherical particles with a size range from 7 to 19 nm. The spectrophotometric and biological activity studies based on Cu-MOF-NPs were analyzed. The results indicated that Cu-MOF-NPs (C1) were successfully used as biosensors for the assessment of the triiodothyronine hormone (T3). The calibration plot was achieved over the concentration range of 40.0-100.0 ng dl-1 T3 with limits of detection (LOD) and quantitation (LOQ) of 1.46 and 4.85 ng dl-1, respectively, and a correlation coefficient (r) of 0.973. Moreover, the Cu-MOF-NPs (C1) show more enhanced biological activity against various pathogens (five strains of bacteria: Gram positive and Gram negative) when compared to an antibacterial agent and the effectiveness of Cu-MOF-NPs increases with increasing particle dose. The interactions of MOF-NPs (C1) with the biological targets were studied.