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Background: The prostate-specific antigen (PSA) is an important cancer biomarker that is commonly utilized in the diagnosis of prostate cancer. The development of a PSA determination technique that is rapid, simple, and inexpensive, in addition to highly accurate, sensitive, and selective, remains a formidable obstacle. Methods: In this study, we developed a practical biosensor based on Zn(II) metal–organic framework nanoparticles (Zn-MOFs-NPs). Many spectroscopic and microanalytical tools are used to determine the structure, morphology, and physicochemical properties of the prepared MOF. Results: According to the results, Zn-MOFs-NPs are sensitive to PSA, selective to an extremely greater extent, and stable in terms of chemical composition. Furthermore, the Zn-MOFs-NPs did not exhibit any interferences from other common analytes that might cause interference. The detection limit for PSA was calculated and was 0.145 fg/mL throughout a wide linear concentration range (0.1 fg/mL–20 pg/mL). Conclusions: Zn-MOFs-NPs were successfully used as a growing biosensor for the monitoring and measurement of PSA in biological real samples. [ABSTRACT FROM AUTHOR]