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  3. Dual-gold nanoprobe system enables highly sensitive SERS detection of glycosylated CEs1 in HepG-2 cell culture supernatant

Dual-gold nanoprobe system enables highly sensitive SERS detection of glycosylated CEs1 in HepG-2 cell culture supernatant

  • Talanta. 2026 Jul 1:304:129561. doi: 10.1016/j.talanta.2026.129561.
Huihui Zeng 1,  Jinru Cao 2,  Yaqin Zhan 1,  Huaqiu Xu 1,  Yue Wang 1,  Ziyu Xu 1,  Tingsha Chen 1,  Yan Li 1,  Wenyi Huang 1,  Hao Cheng 1,  Si Gao 1,  Jun Feng 3
Affiliations

Affiliations

  • 1 Guangxi Key Laboratory of Green Processing of Sugar Resources, Department of Medicine/College of Biological and Chemical Engineering, Guangxi University of Science and Technology, Liuzhou, Guangxi, 545005, China.
  • 2 Dongguan Key Laboratory of Precision Molecular Diagnostics, The Affiliated Dongguan Songshan Lake Central Hospital, Guangdong Medical University, Dongguan, Guangdong, 523200, China.
  • 3 Guangxi Key Laboratory of Green Processing of Sugar Resources, Department of Medicine/College of Biological and Chemical Engineering, Guangxi University of Science and Technology, Liuzhou, Guangxi, 545005, China. Electronic address: [email protected].
PMID: 41722413 DOI: 10.1016/j.talanta.2026.129561
Abstract

Carboxylesterase 1 (CES1) is crucial in liver injury and hepatocellular carcinoma (HCC) progression, necessitating rapid, sensitive detection methods for its glycosylated form. This study developed a surface-enhanced Raman scattering (SERS) biosensor using a dual-gold nanoprobe system for quantifying specifically glycosylated CES1 in HepG-2 Cell Culture supernatant. A glycan probe was constructed from 10 nm AuNPs modified with Raman reporter DTNB and diphenylcyclooctyne (DIBO) to recognize and couple metabolically labelled, glycosylated CES1 via a copper-free click reaction. A protein probe, comprising 40 nm AuNPs conjugated with anti-CEs1, was used for specific capture of glycosylated CES1. The proximity of these two probes induces a spatially controllable SERS effect, generating an enhanced SERS signal. This method achieved quantitative detection of glycosylated CES1 in Cell Culture supernatants, demonstrating excellent linearity from 0.1 nM to 6.0 μM, with a correlation coefficient (R2) of 0.992 and a detection limit of 16 pM. It was successfully applied to monitor glycosylated CES1 release after HepG-2 cell stimulation by hepatotoxic drugs, confirming a positive correlation with both drug concentration and incubation time. This strategy is rapid, sensitive, and highly selective, enabling accurate measurement in complex matrices. It provides a novel approach for early warning of liver injury and HCC and for exploring the biological significance of glycosylated CES1.

Keywords

Carboxylesterase 1; Dual-gold nanoprobes; Glycosylation; Metabolic glycan labeling; Surface-enhanced Raman scattering (SERS).

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