Turn-on fluorometric and colorimetric dual-mode assay based on the in situ formation of silicon nanoparticles for the determination of β-galactosidase

  • Anal Chim Acta. 2025 Sep 1:1365:344260. doi: 10.1016/j.aca.2025.344260.
Sikai Wang  1 Weiping Liu  2 Lei Wang  3 Lin Lv  3 Yuanyuan Zhang  4 Yan Zhao  5
Affiliations
  • 1. School of Science, Xihua University, Chengdu, 610039, China; School of Food and Bioengineering, Xihua University, Chengdu, 610039, China.
  • 2. Department of Transfusion Medicine, Zigong First People's Hospital, Zigong, 643000, Sichuan, China.
  • 3. School of Science, Xihua University, Chengdu, 610039, China.
  • 4. School of Science, Xihua University, Chengdu, 610039, China. Electronic address: [email protected].
  • 5. School of Science, Xihua University, Chengdu, 610039, China; Asymmetric Synthesis and Chiral Technology Key Laboratory of Sichuan Province, Xihua University, Chengdu, 610039, China. Electronic address: [email protected].
Abstract

Background: β-Galactosidase (β-Gal) is widely acknowledged as a crucial biomarker for primary ovarian Cancer. Therefore, precise analysis of β-Gal plays a significant role in the early diagnosis of primary ovarian Cancer. In contrast to organic molecule probes, silicon nanoparticles (Si NPs) have garnered significant attention owing to their outstanding photoluminescence stability and good biocompatibility. Nevertheless, most nanomaterials synthesized under stringent conditions generally produce a single signal that is susceptible to various external influences. Thus, there is an urgent need to develop multi-mode sensing techniques based on Si NPs synthesized under mild conditions for the measurement of β-Gal.

Results: In this study, we developed an innovative and straightforward turn-on fluorometric and colorimetric dual-mode sensing method using silicon nanoparticles (Si NPs) for the detection of β-Gal activity. Such Si NPs, exhibiting a brown-red color and yellow fluorescence, can be readily synthesized under mild conditions by mixing 3-(2-aminoethylamino)propyl-dimethoxymethylsilane and p-aminophenol (AP), both of which are non-fluorescent. By substituting AP with 4-aminophenyl-β-d-galactopyranoside, a substrate that can be hydrolyzed to AP by β-Gal, a sensing platform for detecting β-Gal activity can be constructed. The sensing system is capable of detecting β-Gal within the concentration ranges of 0.1-90 U/L for the fluorometric assay and 0.25-50 U/L as well as 50-100 U/L for the colorimetric assay, respectively. The limits of detection were determined to be 0.052 and 0.171 U/L, accordingly.

Significance: This work introduces, for the first time, a straightforward and convenient dual-mode turn-on fluorometric and colorimetric method utilizing novel Si NPs to determine β-Gal activity. The developed sensing assay was effectively utilized to measure β-Gal levels in human serum and to screen for β-Gal inhibitors, demonstrating its practicality and promising potential in the diagnosis of ovarian Cancer and pharmaceutical development.

Keywords
Dual-mode assay; Inhibitor screening; Silicon nanoparticles; β-Galactosidase.
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