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glypican-3
MICROCHIMICA ACTA · 2024 ·Vol. 191 ·期 11 ·1-17 · IF 5.3

A sandwich-type electrochemical sensor based on RGO-CeO2-Au nanoparticles and double aptamers for ultrasensitive detection of glypican-3

Liang Jintao,Liang Jianlu,Xiao Xinkai,Guo Fei,Zhan Yulian,Zhou Xueqing,Zhou Zhide,Li Guiyin

A sandwich-type electrochemical aptasensor for ultrasensitive detection of glypican-3 (GPC3) was constructed using GPC3 aptamer (GPC3 Apt ) labelled reduced graphene oxide-cerium oxide-gold nanoparticles (RGO-CeO 2 -Au NPs) as the signal probe and the same GPC3 Apt as the capture probe. The electrochemical redox properties of CeO 2 (Ce 3+ /Ce 4+ ) in the RGO-CeO 2 -Au NPs indicate the electrochemical signals. When the target GPC3 was present, an “aptamer-protein-aptamer” sandwich structure was formed on the sensing interface due to the specific binding between the protein and aptamers, resulting in an increased electrochemical redox signal detected by differential pulse voltammetry (DPV) technique. Under optimal conditions, the established aptasensor exhibited a logarithmic linear relationship between the current response and GPC3 concentration in the range 0.001–100.0 ng/mL, with a minimum detection limit of 0.74 pg/mL. Using the spike-recovery tests for measurement of the human serum samples, the recovery was from 99.26 to 114.01%, and the RSD range was 3.04 to 5.34%. Furthermore, the sandwich-type electrochemical aptasensor exhibited excellent performance characteristics such as good stability, high specificity, and high sensitivity, demonstrating effective detection of GPC3 in human serum samples and can be used as a clinical detection tool for GPC3. Graphical Abstract

✓ 已验证 产品关联 DOI PubMed 高影响因子
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MICROCHEMICAL JOURNAL · 2024 · IF 4.9

A highly sensitive sandwich-type electrochemical sensor for detection glypican-3 based on H-rGO-CMC@Pt NPs and aptamers

Xiaohong Tan,Wenzhan Li,Wei He,Tingting Yu,Bingbing Wan,Yong Huang,Jintao Liang,Guiyin Li

Glypican-3 (GPC3), which is a surface heparan sulfate proteoglycan, has been widely known the ideal biomarker for diagnosis and treatment of hepatocellular carcinoma (HCC). Accurate and sensitive detection of serum GPC3 level is an important and challenging task. In this paper, a sandwich electrochemical aptasensor was designed to specifically detect serum GPC3 level through aptamer-target-aptamer recognition. A GPC3 aptamer (GPC3 Apt ) immobilized on the surface of gold nanoparticles@reduced graphene oxide modified screen-printed electrode (Au NPs@rGO/SPCE) was used as the capture probe. Hemin-reduced graphene oxide-carboxymethyl chitosan@platinum nanoparticles (H-rGO-CMC@Pt NPs), with good mimicking peroxidases activity, were labeled with amination GPC3 aptamer, which was employed as the signal probe. Once the target GPC3 was anchored on the surface of GPC3 Apt /Au NPs@rGO/SPCE, the H-rGO-CMC@Pt NPs-GPC3 Apt was further specifically bound to the GPC3, forming the aptamer-target-aptamer sandwich structure, which can catalyze the oxidation of hydroquinone (HQ) to benzoquinone (BQ) and lead the oxidation current of HQ recorded by DPV to changing. The linear relationship between the current signal of oxidation of HQ and GPC3 concentration range of 0.0001–3.0 µg/mL and 3.0–60.0 µg/mL, with a low detection limit (LOD) of 0.0685 ng/mL. Additionally, the electrochemical aptasensor was successfully applied for the detection of GPC3 in human serum samples with the recovery of 99.95–104.06 % and relative standard deviation (RSD) of 1.31–5.22 %. Thus, the sandwich aptasensor could provide a new strategy for detection of serum GPC3 level and improve the early diagnosis rate of HCC.

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ANALYTICAL AND BIOANALYTICAL CHEMISTRY · 2022 ·Vol. 414 ·期 22 ·6441-6453 · IF 4.478

A highly sensitive strategy for glypican-3 detection based on aptamer/gold carbon dots/magnetic graphene oxide nanosheets as fluorescent biosensor

Li Guiyin,Chen Wei,Mi Danhong,Wang Bo,Li HaiMei,Wu Guangxiong,Ding Ping,Liang Jintao,Zhou Zhide

Hepatocellular carcinoma (HCC) is the second leading cause of cancer-related deaths in China. Glypican-3 (GPC3) is a specific antigen related to HCC, which is widely used in clinical detection as a reliable marker of HCC. In this paper, a highly sensitive homogeneous apatasensor was designed for GPC3 detection based on fluorescence resonance energy transfer (FRET) where the GPC3 aptamer labelled gold carbon dots (AuCDs-GPC3 Apt ) are used as a donor and magnetic graphene oxide (Fe 3 O 4 /GO) nanosheets are used as an acceptor. A one-step hydrothermal method was used to synthesize AuCDs to provide sufficient fluorescence. The FRET phenomenon exists between AuCDs-GPC3 Apt and Fe 3 O 4 /GO, which weakens the fluorescence intensity of the whole system. When the target GPC3 is added to the FRET system, the fluorescent AuCDs-GPC3 Apt binds to the GPC3 and forms a folded structure, which leads to AuCDs-GPC3 Apt separation from Fe 3 O 4 /GO nanosheets. The Fe 3 O 4 /GO is then magnetically separated so that the fluorescence of free labelled AuCDs-GPC3 Apt is restored. Under the optimum conditions, the fluorescence recovery rate is linearly correlated with the concentration of GPC3 (5–100 ng·mL −1 ) and the detection limit is 3.01 ng·mL −1 (S/N = 3). This strategy shows recoveries from 98.76 to 101.29% in real human serum samples and provides an immediate and effective detection method for the quantification of GPC3 with great potential applications for early diagnosis of HCC. Graphical abstract A sensitive homogeneous FRET-based apatasensor was designed for GPC3 detection where the AuCDs-GPC3 Apt is a donor and Fe 3 O 4 /GO nanosheets are an acceptor. The GPC3 fluorescent aptasensor combines wider output range with low cost, high specificity, and good anti-interference.

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Journal of Biomedical Nanotechnology · 2021 ·Vol. 17 ·期 4 ·677-690 · IF 4.099

Targeted Delivery of Glypican 3 (GPC3) Antibody-Modified MicroRNA (miR let-7b-5p) Polymer Nanoparticles to Sorafenib-Resistant Hepatsocellular Carcinoma Cells

Zhou Shuping,Ma Yongfang,Liu Xueke,Yu Pan,Huang Ning,Song Li,Xu Ruyue,Huo Zhen,Zhu Tao,Tang Xiaolong

The miR let-7b-5p (a kind of microRNAs) has many pathophysiological regulation effects, including human hepatocellular carcinoma (HCC) pathogenesis. This study investigated whether nanoparticle-mediated miR let-7b-5p could jointly enhance the therapeutic effect of sorafenib on HCC by inhibiting the proliferation of HCC cells, inducing apoptosis, and reversing drug resistance. We evaluated the level of miR let-7b-5p in sorafenib-resistant HepG2 cells (HepG2R) and HepG2 HCC cells by qRT-PCR and analyzed the biological effects of hepatocellular carcinoma treated with sorafenib with miR let-7b-5p, and further studied the toxicity of nanoparticles (Ab-miR-NPs) that deliver miR let-7b-5p mimics and target GPC3 on the surface of hepatocellular carcinoma cells. Results showed that, in HepG2 cells, the expression level of miR let-7b-5p was significantly higher than that in HepG2R cells. Targeted nanoparticle Ab-miR-NPs mediated the delivery of miR let-7b-5p to the HCC cytoplasm and released miRNA after being broken down, down-regulating the expression of IGF1R and inhibiting AKT/mTOR and Ras/Raf signal transmission. Ab-miR-NPs not only enhanced the proliferation of sorafenib in cultured HepG2R cells and induced cell apoptosis efficiency, but they also improved the anti-tumor activity in the mouse models. These results indicated that GPC3 antibody-modified PLGA-PLL (polylactic acid-glycolic acetic copolymer grafted hyper-branched polylysine) loaded miR let-7b-5p polymer nanoparticles combined with sorafenib may be a new treatment strategy for HCC resistant to sorafenib.

✓ 已验证 DOI PubMed 高影响因子
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BIOELECTROCHEMISTRY · 2024 · IF 4.8

A dual-signal output electrochemical aptasensor for glypican-3 ultrasensitive detection based on reduced graphene oxide-cuprous oxide nanozyme catalytic amplification strategy

Guiyin Li,Huafu Feng,Xinhao Li,Shengnan Li,Jintao Liang,Zhide Zhou

Glypican-3 (GPC3) is an essential reference target for hepatocellular carcinoma detection, follow-up and prediction. Herein, a dual-signal electrochemical aptasensor based on reduced graphene oxide-cuprous oxide (RGO-Cu 2 O) nanozyme was developed for GPC3 detection. The RGO-Cu 2 O nanoenzyme displayed excellent electron transport effect, large specific surface area and outstanding peroxidase-like ability. The differential pulse voltammetry (DPV) signal of Cu 2 O oxidation fraction and the chronoamperometry ( i-t ) signal of H 2 O 2 decomposition catalyzed by RGO-Cu 2 O nanozyme were used as dual-signal detection. Under optimal conditions, the log-linear response ranges were 0.1 to 500.0 ng/mL with the limit of detection 0.064 ng/mL for DPV technique, and 0.1–50.0 ng/mL for i-t technique (detection limit of 0.0177 ng/mL). The electrochemical aptasensor has remarkably analytical performance with wide response range, low detection limit, excellent repeatability and specificity, good recovery in human serum samples. The two output signals of one sample achieve self-calibration of the results, effectively avoiding the occurrence of possible leakage and misdiagnosis of a single detection signal, suggesting that it will be a promising method in the early biomarker detection.

✓ 已验证 DOI PubMed 高影响因子
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ELECTROCHIMICA ACTA · 2022 · IF 7.336

Label-free detection of glypican-3 using reduced graphene oxide /polyetherimide/gold nanoparticles enhanced aptamer specific sensing interface on light-addressable potentiometric sensor

Guiyin Li,Bo Wang,Le Zhao,Xiaohang Shi,Guanxiong Wu,Wei Chen,Lixian Sun,Jintao Liang,Zhide Zhou

Glypican-3 (GPC3) protein is considered as an effective and specific biomarker for the accurate diagnosis of hepatocellular carcinoma (HCC), and there is a critical demand to construct strategies with shorter time, lower cost noise and higher sensitivity. In this study, a biosensor for label-free detection of GPC3 was prepared by modifying GPC3 aptamer (GPC3 Apt ) on the surface of Si-based light addressable potentiometric sensor (LAPS) chip modified by reduced graphene oxide/polyetherimide/gold nanoparticles (rGO/PEI/AuNPs). Among them, GPC3 Apt with a low equilibrium dissociation constant was used as a biorecognition molecule to recognize and capture GPC3. The rGO/PEI/AuNPs decorated on the LAPS surface could enlarge the surface area for GPC3 Apt immobilization and provide more active sites for GPC3 Apt to bind effectively with the target GPC3. When a bias voltage was provided to the LAPS system, the forming GPC3-GPC3 Apt complexs on the sensing interface of LAPS would cause the potential of the sensitive membrane surface to change and generate a corresponding offset response in photocurrent-voltage (I-V) curves. The potential shift and GPC3 concentration showed a good linear range from 0.1 to 100.0 μg/mL with a detection limit of 40.0 ng/mL (S/ N  = 3) and a sensitivity of 1.33638 mV/μg/mL. In addition, the LAPS detection system also has good specificity and stability, which can be used for the detection of GPC3 in spiked serum samples.

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Journal of Biomedical Nanotechnology · 2021 ·Vol. 17 ·期 12 ·2444-2454 · IF 4.099

Highly Sensitive Electrochemical Aptasensor for Detection of Glypican-3 Using Hemin-Reduced Graphene Oxide-Platinum Nanoparticles Coupled with Conductive Reduced Graphene Oxide-Gold Nanoparticles

Li Guiyin,Li HaiMei,Chen Wei,Chen Huijiang,Wu Guanxiong,Tan Mingxiong,Liang Jintao,Zhou Zhide

An electrochemical aptasensor for quantitatively detecting glypican-3 (GPC3) was constructed by combining hemin-reduced graphene oxide-platinum (H-rGO-Pt) nanoparticles (NPs) with reduced graphene oxide-gold (rGO-Au) nanoparticles (NPs). Herein, the rGO-Au NPs deposited onto screen-printed electrodes resulted in signal amplification due to their large surface areas. Meanwhile, highly conductive H-rGO-Pt NPs acted as a sensing medium that improved electrical conductivity and as an indicator for monitoring peak current for determination. A GPC3 aptamer (GPC3apt) with a low equilibrium dissociation constant was used as a bio-recognition molecule. GPC3apt specifically captured GPC3 proteins and formed aptamer-GPC3 complexes, which impeded electron transfer and thus hampered the redox signal of hemin in H-rGO-Pt NPs. This developed electrochemical aptasensor showed a linear response to GPC3 (from 0.001 μg/mL to 10 μg/mL) and had a detection limit of 0.001 μg/mL. This work provides a low-cost and highly sensitive detection with and good recovery for GPC3 and holds great promise for the clinical diagnosis of hepatocellular carcinoma.

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Biomaterials Science · 2022 ·Vol. 10 ·期 23 ·6804-6817 · IF 7.59

A label-free electrochemical aptasensor based on platinum@palladium nanoparticles decorated with hemin-reduced graphene oxide as a signal amplifier for glypican-3 determination

Guiyin Li,Wenzhan Li,Shengnan Li,Xinhao Li,Xiaoqing Yao,Wen Xue,Jintao Liang,Jiejing Chen,Zhide Zhou

Glypican-3 (GPC3) is a membrane-associated proteoglycan that is specifically upregulated in hepatocellular carcinoma (HCC) and has become one of the most promising biomarkers closely related to the occurrence and development of HCC. In this work, platinum@palladium nanoparticles decorated with hemin-reduced graphene oxide (H-rGO-Pt@Pd NPs) were used not only as a support for GPC3 aptamer (GPC3Apt) immobilization, but also as a new redox nanoprobe in electrochemical analysis for the determination of GPC3. The electrochemical aptasensor involved a reaction cell with a three-electrode system, and the differential pulse voltammetry (DPV) technique was adopted. In the presence of GPC3, the formed GPC3Apt–GPC3 complexes had stable structures and were cleaved from the electrode surface, leading to more electroactive H-rGO-Pt@Pd NPs repelling freely from the GPC3Apt/H-rGO-Pt@Pd NPs and thus to the increase of the oxidation peak current of hemin in H-rGO-Pt@Pd NPs. Under optimal conditions and a working voltage of +700 mV (vs. Ag/AgCl), the label-free electrochemical GPC3 aptasensor showed superior performance with a wider concentration linear range (0.001–10.0 μg mL−1), a lower limit of detection (LOD) (0.181 ng mL−1, S/N = 3), a higher sensitivity (0.0446 μA μM−1 cm−2) and good selectivity. Furthermore, the fabricated aptasensor was applied to GPC3 determination in human serum samples with satisfactory recoveries of 94.3%–119% and RSDs of 0.15%–5.78%. The current work provides a flexible approach for the rapid and sensitive analysis of GPC3 and has a broad application prospect in the diagnosis of HCC.

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