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It is crucial to detect Pb2+ accurately and rapidly. This work proposes an ultra-sensitive optical fiber surface plasmon resonance (SPR) sensor functionalized with glutathione (GSH) for label-free detection of the ultra-low Pb2+ concentration, in which the refractive index (RI) sensitivity of the multimode-singlemode-multimode (MSM) hetero-core fiber is largely enhanced by the gold nanoparticles (AuNPs)/Au film coupling SPR effect. The GSH is modified on the fiber as the sensing probe to capture and identify Pb2+ specifically. Its working principle is that the Pb2+ chemically reacts with deprotonated carboxyl groups in GSH through ligand bonding, resulting in the formation of stable and specific chelates, inducing the variation of the local RI on the sensor surface, which in turn leads to the SPR wavelength shift in the transmission spectrum. Attributing to the AuNPs, both the Au substrates can be fully functionalized with the GSH molecules as the probes, which largely increases the number of active sites for Pb2+ trapping. Combined with the SPR effect, the sensor achieves a sensitivity of 2.32 × 1011 nm/M and a limit of detection (LOD) of 0.43 pM. It also demonstrates exceptional specificity, stability, and reproducibility, making it suitable for various applications in water pollution, biomedicine, and food safety. [ABSTRACT FROM AUTHOR]