A novel sensing platform for the determination of alkaline phosphatase based on SERS-fluorescent dual-mode signals

Alkaline phosphatase (ALP), as an important biomarker, is closely associated with various diseases. Multi-mode sensing platforms can combine the advantages of different technologies and solve their inherent or practical limitations. Herein, we developed a sensing platform for the determination of alkaline phosphatase (ALP) in human serum based on SERS-fluorescent dual-mode assay. Based on the fact that ALP can trigger the in-situ reaction between o-phenylenediamine (OPD) and ascorbic acid (AA), we connected gold nanoparticles (AuNPs) to 3,4-diaminobenzene-thiol (OPD(SH)) through an Au–S covalent bond to synthesize a nanoprobe (OPD(S)-AuNPs). The nanoprobe provides a unique interactive ammonium group for the diol group of AA, which was then used to generate an N-heterocyclic compound that can exhibit good SERS and fluorescence signals without adding SERS reporter and fluorophores or quantum dots (QDs). When being excited at different wavelengths as 360 nm and 785 nm, the fluorescence and SERS signals can be separately generated, which can avoid the disturbance from each other. The response of the fluorescence system was linear from 1.0 to 20 mU mL⁻¹ (R² = 0.994) with a detection limit of 0.3 mU mL⁻¹, while that of the SERS system was linear from 0.5 to 10 mU mL⁻¹ (R² = 0.998) with a detection limit of 0.2 mU mL⁻¹. The sensing platform developed was further employed in ALP inhibitor evaluation.